DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 12/5/2025 has been entered.
Election/Restrictions
Previously withdrawn claims 29-30 are rejoined in view of the art.
Status of Claims
Claims 1-3, 5, 7-8, 10-15, 17-19, 21-22, 24-32 are under examination on the merits.
Claim Rejections - 35 USC § 112
Written Description
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-3, 5, 7-8, 10-15, 17-19, 21-22 & 24-32 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection set forth in the Office action mailed 10/9/2025, as applied to claims 1-5, 7-8, 10-19, 21-22, 24-28 & 31-32. Applicant's arguments filed 11/14/2025 have been fully considered but they are not persuasive.
Claims 1-3, 5, 7-8, 10-15, 17-19, 21-22, 24-32 are broadly drawn to a modified maize plant comprising reduced expression or activity of an endogenous CYP710A gene wherein the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 and methods for producing such a plant or producing a commodity plant product from such a plant or a commodity plant product produced. Claims 2 & 14 require that the CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2 and claims 3 & 15 require that the endogenous CYP710A gene comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1. Claim 10 requires that the modified maize plant comprises a CYP710A gene having at least 80% identity to SEQ ID NO: 3; however, the CYP710A gene of claim 10 is not required to be endogenous to a maize plant.
An endogenous CYP710A gene encompasses any maize CYP710A gene, including any and all paralogs. Although the claims require reduced expression or activity of an endogenous CYP710A gene and one or more nucleotide modifications at a genomic locus comprising the CYP710A gene, the claims do not require that the nucleotide modifications in the CYP710A gene result in the reduced expression or activity. Reduced expression or activity of an endogenous CYP710A gene encompasses genetic modifications to this gene, including its coding sequence, intronic regions, or regulatory sequences. Reduced expression or activity also encompasses genetic modifications to other endogenous genes that regulate CYP710A or the introduction of exogenous elements that reduce expression or activity. “Modified” additionally encompasses epigenetic modifications to the CYP710A gene or associated genetic elements.
A polypeptide comprising an amino acid sequence having at least 95% or 99% sequence identity to the 513 amino acid-long SEQ ID NO: 2 encompasses sequences with 25 or 5 amino acid substitutions relative to SEQ ID NO: 2. Nucleotide sequences with 95% identity to the 1542 nucleotide long SEQ ID NO:1 would have 77 nucleotide substitutions relative to SEQ ID NO: 1.
A Sterol C-22 desaturase-like protein in maize is known in the art with 95.3% sequence identity to SEQ ID NO: 2. See alignment below. This sequence, SEQ ID NO: 66893 taught by Wu et al US 2012/0017338A1 (1/19/2012), has only 1 mismatch and one conservative amino acid substitution relative to instant SEQ ID NO: 2. The remaining sequence divergence is due to indels. NCBI Reference Sequence NP_001307723.1 (available 6/27/2021) also teaches an endogenous maize Cytochrome P450 710A1-like protein located on chromosome 3, but this sequence also has 99% sequence identity to instant SEQ ID NO: 2, with only substitutions and at the same two amino acids as Wu’s SEQ ID NO: 66893. See second alignment below. Finally, the art describes maize DNA sequences with as little as 90% sequence identity to instant SEQ ID NO: 1 (Wu et al US 2012/0017338A1 SEQ ID NO: 25114, see third alignment below), but this single sequence with only 7 nucleotide mismatches does not describe the full scope of claimed variation in endogenous maize CYP710A genes. Endogenous maize CYP710A genes, which encode sterol C-22 desaturases with as little as 95% sequence identity to SEQ ID NO: 2 with sequence variation across the full encoded polypeptide are not described in the art.
US-12-812-263A-66893
Sequence 66893, US/12812263A
Publication No. US20120017338A1
GENERAL INFORMATION
APPLICANT: Kovalic, David
APPLICANT: Wu, Wei et al
APPLICANT: Qiu, Bo-Xing
APPLICANT: Tabaska, Jack E
TITLE OF INVENTION: ISOLATED NOVEL NUCLEIC ACID AND PROTEIN MOLECULES FROM CORN AND
TITLE OF INVENTION: METHODS OF USING THOSE MOLECULES TO GENERATE TRASGENIC PLANT
TITLE OF INVENTION: WITH ENHANCED AGRONOMIC TRAITS
FILE REFERENCE: 38-21(55305)B-0001
CURRENT APPLICATION NUMBER: US/12/812,263A
CURRENT FILING DATE: 2011-06-24
PRIOR APPLICATION NUMBER: PCT/09/00153
PRIOR FILING DATE: 2009-01-09
PRIOR APPLICATION NUMBER: 61/011,138
PRIOR FILING DATE: 2008-01-15
NUMBER OF SEQ ID NOS: 83558
SEQ ID NO 66893
LENGTH: 499
TYPE: PRT
ORGANISM: Zea mays
FEATURE:
OTHER INFORMATION: Coding DNA Sequence Number=SeqID_25114
FEATURE:
OTHER INFORMATION: Gene Ontology: Mol. function=monooxygenase activity; Cellular
component=integral to membrane; Biological process=electron
transport
FEATURE:
OTHER INFORMATION: Homolog annotation: Hit_ID=Q9LDQ4; Match level="QueryCoverage=93\%,
HitCoverage=94\%, E-value=0.0, Identity=84\%"; Hit
description=Sterol C-22 desaturase-like
FEATURE:
OTHER INFORMATION: Pfam annotation: Domain name=p450; Domain location=48-489; Score
=10.1; E_value=2e-11; Accession=PF00067.12;
Description=Cytochrome P450
Query Match 95.3%; Score 2569; Length 499;
Best Local Similarity 95.7%;
Matches 494; Conservative 1; Mismatches 1; Indels 20; Gaps 2;
Qy 1 MAAQSLSVPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
|||||||||||: |||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MAAQSLSVPVPMPSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
Qy 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
Qy 121 HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRS- 179
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG 180
Qy 180 --GGGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFA 237
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GGGGGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFA 240
Qy 238 FRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAH 297
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 FRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAH 300
Qy 298 TDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTA 357
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 TDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTA 360
Qy 358 ETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGF 417
||||||||||||||||||||||||||||||||||||||||| ||
Db 361 ETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYT-----------------GF 403
Qy 418 PDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFR 477
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 404 PDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFR 463
Qy 478 RERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 513
||||||||||||||||||||||||||||||||||||
Db 464 RERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 499
Score
Expect
Method
Identities
Positives
Gaps
1036 bits(2678)
0.0
Compositional matrix adjust.
511/516(99%)
512/516(99%)
3/516(0%)
Query 1 MAAQSLSVPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
MAAQSLSVPVP+ SSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV
Sbjct 1 MAAQSLSVPVPMPSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
Query 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG
Sbjct 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
Query 121 HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG 180
HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG
Sbjct 121 HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG 180
Query 181 GG---AFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFA 237
GG AFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFA
Sbjct 181 GGGGGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFA 240
Query 238 FRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAH 297
FRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAH
Sbjct 241 FRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAH 300
Query 298 TDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTA 357
TDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTA
Sbjct 301 TDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTA 360
Query 358 ETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGF 417
ETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGF
Sbjct 361 ETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGF 420
Query 418 PDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFR 477
PDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFR
Sbjct 421 PDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFR 480
Query 478 RERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 513
RERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF
Sbjct 481 RERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 516
US-12-812-263A-25114
(NOTE: this sequence has 1 duplicate in the database searched)
Sequence 25114, US/12812263A
Publication No. US20120017338A1
GENERAL INFORMATION
APPLICANT: Kovalic, David
APPLICANT: Wu, Wei et al
APPLICANT: Qiu, Bo-Xing
APPLICANT: Tabaska, Jack E
TITLE OF INVENTION: ISOLATED NOVEL NUCLEIC ACID AND PROTEIN MOLECULES FROM CORN AND
TITLE OF INVENTION: METHODS OF USING THOSE MOLECULES TO GENERATE TRASGENIC PLANT
TITLE OF INVENTION: WITH ENHANCED AGRONOMIC TRAITS
FILE REFERENCE: 38-21(55305)B-0001
CURRENT APPLICATION NUMBER: US/12/812,263A
CURRENT FILING DATE: 2011-06-24
PRIOR APPLICATION NUMBER: PCT/09/00153
PRIOR FILING DATE: 2009-01-09
PRIOR APPLICATION NUMBER: 61/011,138
PRIOR FILING DATE: 2008-01-15
NUMBER OF SEQ ID NOS: 83558
SEQ ID NO 25114
LENGTH: 1500
TYPE: DNA
ORGANISM: Zea mays
FEATURE:
OTHER INFORMATION: Peptide Sequence Number=SeqID_66893
Query Match 90.8%; Score 1399.8; Length 1500;
Best Local Similarity 95.7%;
Matches 1484; Conservative 0; Mismatches 7; Indels 60; Gaps 2;
Qy 1 ATGGCGGCGCAGTCACTGTCCGTGCCCGTGCCCGTGCTCAGCTCCTTGGACCTCCGCGCG 60
|||||||||||||| |||||||||||||||||| ||| ||||||||||||||||||||||
Db 1 ATGGCGGCGCAGTCTCTGTCCGTGCCCGTGCCCATGCCCAGCTCCTTGGACCTCCGCGCG 60
Qy 61 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCCTTGTACTTCCTGATGGAGCAGGTGTCCTAC 120
||||||||||||||||||||||||||||| ||||||||||||||||||||||||||||||
Db 61 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCTTTGTACTTCCTGATGGAGCAGGTGTCCTAC 120
Qy 121 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTG 180
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTA 180
Qy 181 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 240
Qy 241 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 300
Qy 301 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 360
Qy 361 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 420
Qy 421 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 480
Qy 481 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 540
Qy 541 ---------GGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACC 591
|||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 GGCGGCGGTGGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACC 600
Qy 592 TCGCAGACGGTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGG 651
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 TCGCAGACGGTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGG 660
Qy 652 GACTACAACCTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCG 711
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 GACTACAACCTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCG 720
Qy 712 TTCCGGCGCGCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGG 771
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TTCCGGCGCGCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGG 780
Qy 772 CAGAGCAAGGCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATG 831
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 CAGAGCAAGGCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATG 840
Qy 832 CAGGACACGCTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCAC 891
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 CAGGACACGCTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCAC 900
Qy 892 ACCGACGACGAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCG 951
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 ACCGACGACGAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCG 960
Qy 952 TCCACCTCGTCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCC 1011
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 TCCACCTCGTCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCC 1020
Qy 1012 CGCGTGCGCGCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCG 1071
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 CGCGTGCGCGCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCG 1080
Qy 1072 GAGACGATCCAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGG 1131
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 GAGACGATCCAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGG 1140
Qy 1132 CCCCCCGCGACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTAC 1191
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 CCCCCCGCGACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTAC 1200
Qy 1192 ACGGTGCCCAAGGGCGCCATCGTGTTCCCGTCCGTGTACGAGTCGTCGTTCCAGGGGTTC 1251
|| |||||||
Db 1201 AC---------------------------------------------------GGGGTTC 1209
Qy 1252 CCGGACGCGGAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTG 1311
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1210 CCGGACGCGGAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTG 1269
Qy 1312 GCGTACAGGCGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAGCGG 1371
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1270 GCGTACAGGCGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAGCGG 1329
Qy 1372 TACGCGCTGAACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTGGACTTCCGG 1431
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1330 TACGCGCTGAACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTGGACTTCCGG 1389
Qy 1432 CGGGAGAGGACGCCCGGGTGTGATGACCCGGTGTACATGCCCACCATCGTGCCCAAGGAC 1491
|||||||||||||||||||| || ||||||||||||||||||||||||||||||||||||
Db 1390 CGGGAGAGGACGCCCGGGTGCGACGACCCGGTGTACATGCCCACCATCGTGCCCAAGGAC 1449
Qy 1492 GGCTGCTCCGTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 1542
|||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1450 GGCTGCTCCGTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 1500
The specification provides the names of 23 plant homologs of CYP710A in 13 species (figure 1C, paragraph [0239]), but the claims are drawn to an endogenous gene in a maize plant. The specification describes a CYP710A gene in maize as SEQ ID NO:1, which encodes SEQ ID NO:2. The specification also provides the nucleotide sequence SEQ ID NO: 3 which is not described but appears to also encode the CYP710A protein and has 90.4% sequence similarity to SEQ ID NO: 1. See alignment below. The specification describes two genetic modifications of the CYP710A gene leading to reduced expression or activity: insertional mutation within the coding sequence and in the 3’UTR. The claims do not require that the nucleotide modifications in the endogenous gene result in the reduced expression or activity; however, given the paucity of nucleotide and amino acid variation described in the art for endogenous maize CYP710A genes, one of ordinary skill in the art would not have understood which nucleotide modifications would have resulted in reduced expression or activity, if any. SEQ ID NOs: 1-2 do not describe variants of the CYP710A gene that are inherently non-functional. Additionally, claims 1, 2, 3, 5, 10-15, 17-19 & 24-32 still encompass reduced expression or activity of the CYP710A gene due to genetic or epigenetic modifications to non-coding regions, regulatory elements, and genes that regulate the endogenous CYP710A gene. Thus, the specification does not describe species over the full scope of claimed endogenous maize CYP710A genes comprising one or more nucleotide modifications nor does the specification describe modifications leading to reduced expression or activity.
US-18-501-470-3
Sequence 3, US/18501470
Publication No. US20240164267A1
GENERAL INFORMATION
APPLICANT: Iowa State University Research Foundation, Inc. (en)
TITLE OF INVENTION: GENETIC MECHANISM TO ENHANCE HEALTHY STEROLS AND HARD ENDOSPERM IN SEEDS (en)
FILE REFERENCE: P14104US01
CURRENT APPLICATION NUMBER: US/18/501,470
CURRENT FILING DATE: 2023-11-03
NUMBER OF SEQ ID NOS: 3
SEQ ID NO 3
LENGTH: 1680
TYPE: DNA
FEATURE:
NAME/KEY: source
LOCATION: 1..1680
QUALIFIERS: mol_type = other DNA
organism = synthetic construct
FEATURE:
NAME/KEY: misc_feature
LOCATION: 1436
QUALIFIERS: note = n indicates an unknown length of sequence present
within the transposon that was not determined
Query Match 90.4%; Score 1394; Length 1680;
Best Local Similarity 91.8%;
Matches 1542; Conservative 0; Mismatches 0; Indels 138; Gaps 1;
Qy 1 ATGGCGGCGCAGTCACTGTCCGTGCCCGTGCCCGTGCTCAGCTCCTTGGACCTCCGCGCG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ATGGCGGCGCAGTCACTGTCCGTGCCCGTGCCCGTGCTCAGCTCCTTGGACCTCCGCGCG 60
Qy 61 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCCTTGTACTTCCTGATGGAGCAGGTGTCCTAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCCTTGTACTTCCTGATGGAGCAGGTGTCCTAC 120
Qy 121 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTG 180
Qy 181 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 240
Qy 241 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 300
Qy 301 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 360
Qy 361 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 420
Qy 421 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 480
Qy 481 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 540
Qy 541 GGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACCTCGCAGACG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 GGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACCTCGCAGACG 600
Qy 601 GTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGGGACTACAAC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGGGACTACAAC 660
Qy 661 CTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCGTTCCGGCGC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 CTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCGTTCCGGCGC 720
Qy 721 GCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGGCAGAGCAAG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 GCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGGCAGAGCAAG 780
Qy 781 GCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATGCAGGACACG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATGCAGGACACG 840
Qy 841 CTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCACACCGACGAC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 CTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCACACCGACGAC 900
Qy 901 GAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCGTCCACCTCG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 GAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCGTCCACCTCG 960
Qy 961 TCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCCCGCGTGCGC 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 TCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCCCGCGTGCGC 1020
Qy 1021 GCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCGGAGACGATC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 GCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCGGAGACGATC 1080
Qy 1081 CAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGGCCCCCCGCG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 CAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGGCCCCCCGCG 1140
Qy 1141 ACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTACACGGTGCCC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 ACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTACACGGTGCCC 1200
Qy 1201 AAGGGCGCCATCGTGTTCCCGTCCGTGTACGAGTCGTCGTTCCAGGGGTTCCCGGACGCG 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 AAGGGCGCCATCGTGTTCCCGTCCGTGTACGAGTCGTCGTTCCAGGGGTTCCCGGACGCG 1260
Qy 1261 GAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTGGCGTACAGG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 GAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTGGCGTACAGG 1320
Qy 1321 CGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAG------------ 1368
||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 CGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAGCGGTACGCGGAG 1380
Qy 1369 ------------------------------------------------------------ 1368
Db 1381 ATAATTGTCATTATAGACGAAGAGCGGACGGGATTCGACGAAATAGAGGCGATGGNGAGA 1440
Qy 1369 ------------------------------------------------------------ 1368
Db 1441 AGCCAACGCCATCGCCTCCATTTCGTCGAATCCCGTCCGCTCTTCGTCTATAATGGCAAT 1500
Qy 1369 ------CGGTACGCGCTGAACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTG 1422
||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 TATCTCCGGTACGCGCTGAACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTG 1560
Qy 1423 GACTTCCGGCGGGAGAGGACGCCCGGGTGTGATGACCCGGTGTACATGCCCACCATCGTG 1482
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1561 GACTTCCGGCGGGAGAGGACGCCCGGGTGTGATGACCCGGTGTACATGCCCACCATCGTG 1620
Qy 1483 CCCAAGGACGGCTGCTCCGTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 1542
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1621 CCCAAGGACGGCTGCTCCGTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 1680
The instant specification describes the maize CYP710A8 protein has having conserved sites and characteristic motifs for CYP710A (paragraph [0238]), but the identity of those motifs and conserved sites is not provided in the instant specification.
CYP710A proteins are known in the art. Morikawa et al (2009) Planta. 229: 1311-1322 (published 3/22/2009, hereafter Morikawa 2009) describes CYP710A proteins has having a conserved F(L/M)FA(A/S)QDA(S/T)(S/T)S sequence at the substrate recognition site on an I helix region, which is crucial for C22-desaturation activity (page 1320, left column, paragraph 1). Morikawa also teaches that the structure of CYP710A genes can vary across species, with introns present in C. reinhardtii and P. patens homologs but absent from Arabidopsis and rice homologs (page 1320, left column, paragraph 1). Fang et al (2024) Genome Biol. Evol. 16(11) published 11/7/2024, after the effective filing date of the instant application; hereafter Fang) teaches motifs of cytochrome P450 proteins that have been lost in CYP710 proteins in some taxa, including a heme-binding region FXXGXRXCXG motif, a C-terminal helix, and a proline-enriched (P/I)PGPX(P/G)XP motif (figure 4; page 11, left column, paragraph 3-right column, paragraph 3).
Although not specifically described in maize, modifications to the expression of endogenous CYP710A genes in plants can result in different phenotypes compared to modifications to the activity of CYP710A proteins. Wang et al (2012) Plant Physiology. 158: 1789-1802 (published April 2012, hereafter Wang) discloses an Arabidopsis Atcyp710A1 T-DNA insertion mutant within the coding sequence that was a null mutant for AtCYP710A1 (page 1797, right column, paragraph 2). Wang describes that Atcyp710A1 mutants with T-DNA insertions upstream of the AtCYP710A1 start codon had a different phenotype with regard to bacterial susceptibility (page 1799, right column, paragraph 2) than those mutants with insertions within the coding region. Modification to expression of a putative CYP710A gene regulatory element led to a different phenotype compared to modification to activity of a CYP710A gene, suggesting that function of CYP710A regulatory elements is also not fully described in the art.
CYP710A genes are diverse in sequence, in motif presence, and in intron structure across species. Maize CYP710A genes that have been described, however, are high in sequence similarity. Examples with as little as 95% sequence similarity to SEQ ID NO: 1 or SEQ ID NO: 2 due to substitutions or across the full sequence are not provided by the art or instant specification. The instant specification fails to describe which modifications to motifs, domains, splicing, and regulatory elements are necessary and sufficient for reduced expression or activity of a CYP710A gene of the present invention to result in increased sitosterol or hard endosperm. The instant specification also fails to describe maize plants comprising endogenous CYP710A genes over the full scope of the claimed genus of CYP710A genes.
Because maize plants comprising endogenous CYP710A genes encoding an amino acid sequence with 95% identity to SEQ ID NO: 2, or with 99% identity to SEQ ID NO: 2 that comprise substitution at a location other than at the 12th and 13th position, or even with a nucleotide sequence of at least 95% sequence identity to SEQ ID NO: 1, nor CYP710A regulatory elements, genes regulating CYP710A, and the genetic and epigenetic modifications thereto that might result in reduced expression or activity and increased sitosterol or hard endosperm, are not described over the full scope of the claims, the method of reducing expression or activity of a CYP710A gene is likewise not described, and the specification fails to provide an adequate written description of the claimed invention. Therefore, given the lack of written description in the specification with regard to the structural and functional characteristics of the compositions used in the claimed methods, Applicant does not appear to have been in possession of the claimed genus at the time this application was filed.
Applicant urges that the amended claims recite the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, which is more narrowly tailored to the working examples in the specification and encompass only sequences having high sequence similarity to SEQ ID NO: 2. Applicant urges that claims 1 & 13 have been amended to specify that the modification is within the CYP710A gene itself (Remarks, page 7, paragraph 3-page 8, paragraph 1).
This argument is unpersuasive, because the claims recite an endogenous maize CYP710A gene encoding a sequence with at least 95% sequence identity to SEQ ID NO: 2, but the instant specification and the prior art does not describe maize plants comprising endogenous sequences over the full scope of the claims. Only one CYP710A maize gene encoding a polypeptide with as little as 95% sequence identity to SEQ ID NO: 2 is described in the art, and the sequence identity was only this divergent because of indels. Endogenous maize CYP710A genes encoding a sequence with 25 or even 5 amino acid substitutions are not known in the art or described by the instant application, so one of ordinary skill in the art would not conclude that Applicant was in possession of maize plants comprising this broadly claimed genus of endogenous maize CYP710A genes at the time of filing.
Additionally of note, the claims do not require that the decrease in activity or expression of the endogenous CYP710A gene is a result of the nucleotide modifications at the CYP710A locus. For example, claims 7 and 8 recite the modified maize plant also comprising a silencing element and claims 21 and 22 specify that the expression or activity is reduced by introducing the silencing element even though nucleotide modifications at the genomic locus are already present.
Scope of Enablement
Claims 1-3, 5, 7-8, 10-15, 17-19, 21-22, 24-32 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a modified plant comprising increased sitosterol or hard endosperm by reducing expression or activity of an endogenous maize CYP710A gene where the CYP710A coding sequence or 3’ UTR is mutated, does not reasonably provide enablement for a modified plant comprising increased sitosterol by reducing expression of any maize endogenous CYP710A gene via nucleotide or epigenetic modifications in any regulatory element operably linked to the gene or an enhancer motif present within a regulatory region of the gene, or by generating alternative spliced variants of the gene or reducing expression or transcriptional activity of encoded polypeptide.
Additionally, the specification, while being enabling for producing a modified plant comprising increased sitosterol or hard endosperm by reducing expression or activity of an endogenous maize CYP710A gene wherein the CYP710A gene has a sequence of SEQ ID NO: 1 and encodes an amino acid sequence of SEQ ID NO: 2, does not reasonably provide enablement for producing a plant with a CYP710A gene with as little as 95% sequence identity to SEQ ID NO: 1 or encoding an amino acid sequence with 95% identity to SEQ ID NO: 2. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make or use the invention commensurate in scope with these claims.
Due to Applicant' s amendment of the claims, the rejection is modified from the rejection set forth in the Office action mailed 10/9/2025, as applied to claims 1-5, 7-8, 10-19, 21-22, 24-28 & 31-32. Applicant's arguments filed 11/14/2025 have been fully considered but they are not persuasive.
A. Claims 1-3, 10-15, 17-18, 24 -28, & 31-32 are drawn to a modified maize plant or progeny or part or cell with increased sitosterol or hard endosperm comprising reduced expression or activity of an endogenous maize CYP710A gene and a method for producing a maize plant with increased sitosterol or hard endosperm comprising reducing expression or activity of an endogenous CYP710A gene in the plant. Claims 29-30 are drawn to a commodity plant product prepared from the plant, plant part, or plant cell. This rejection is regarding reduced expression or activity in the endogenous maize gene leading to the required phenotype.
The claims require reduced expression or activity of an endogenous CYP710A gene and one or more nucleotide modifications at a genomic locus comprising the CYP710A gene. The claims do not require that the nucleotide modifications result in the reduced expression or activity.
The instant specification describes two maize lines with mutations in an endogenous CYP710A gene leading to increased sitosterol (figure 5a; paragraph [0245-0247]; figure 9). The instant specification does not describe the regulatory elements operably linked to the maize CYP710A gene and describes no enhancer motifs. The instant specification does not describe the transcriptional activity or CYP710A expression of the maize CYP710A mutants. The instant specification does not provide examples of splice variants. Thus, the specification does not teach modifications in regulatory regions that would reduce expression or activity of the endogenous CYP710A gene.
The prior art on modifications of CYP710A genes in species other than maize suggests that mutations of regulatory elements may lead to unpredictable results. Gutierrez (2019) Unveiling the Biological Role of Stigmasterol Biosynthesis in Tomato Plants. [Doctoral dissertation, University of Barcelona] (hereafter, Gutierrez) describes a transgenic tomato line for silencing C22DES (a tomato CYP710 homolog: page 24, paragraph 1), and reports that changing transcript levels of C22DES did not lead to a change in stigmasterol content in tomato (page 110, paragraph 2 & page 92, paragraph 2). The C22DES silencing lines thus had a different phenotype than the C22des knockout lines described by Gutierrez (page 99, paragraph 5; table 1). In addition, Gutierrez describes C22DES overexpression lines that have increased sitosterol relative to wildtype (page 94, figure 5b). Wang et al (2012) Plant Physiology. 158: 1789-1802 (published April 2012, hereafter Wang) discloses an Atcyp710A1 T-DNA insertion mutant within the coding sequence that was a null mutant for AtCYP710A1 (page 1797, right column, paragraph 2) and also discloses that T-DNA insertion in the 5’ untranslated region of the Arabidopsis did not have a complete loss of transcripts (page 1797, right column, paragraph 2). Wang describes that Atcyp710A1 mutants with T-DNA insertions upstream of the AtCYP710A1 start codon had a different phenotype with regard to bacterial susceptibility (page 1799, right column, paragraph 2) than mutants with insertions within the coding region. The prior art is silent regarding the phenotype of maize plants with mutations in regulatory regions of CYP710A.
Given that mutations in regulatory regions of a CYP710A gene or modifications that reduce transcription of a CYP710A gene produce different phenotypes and sitosterol profiles compared to mutations that compromise the function of the CYP710A protein, and given that enhancer motifs and alternate splice variants of endogenous CYP710A proteins in maize are undescribed, one of ordinary skill in the art would be required to conduct undue experimentation to identify which mutations or levels of transcription confer the claimed phenotype. Given the unpredictability of the art, claims 1-3, 10-15, 17-18, 24 -28, & 31-32 are not enabled over the full scope of the claimed invention.
Applicant urges that the amended claims recite the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, which is more narrowly tailored to the working examples in the specification and encompass only sequences having high sequence similarity to SEQ ID NO: 2. Applicant urges that claims 1 & 13 have been amended to specify that the modification is within the CYP710A gene itself (Remarks, page 7, paragraph 3-page 8, paragraph 1).
This argument is unpersuasive, because the claims do not require that the decrease in activity or expression of the endogenous CYP710A gene is a result of the nucleotide modifications at the CYP710A locus. Claims 7 and 8 recite the modified maize plant also comprising a silencing element and claims 21 and 22 specify that the expression or activity is reduced by introducing the silencing element even though nucleotide modifications at the genomic locus are already present, which suggests that claims 1 & 13 should not be read as though modifications at the genomic locus are responsible for decrease in activity or expression. The claims encompass modified maize plants wherein the decrease in activity or expression in CYP710A is the result of modification at other regulatory loci, even though the regulatory loci are not taught in the instant disclosure.
Furthermore, given that mutations in regulatory regions of a CYP710A gene or modifications that reduce transcription of a CYP710A gene produce different phenotypes and sitosterol profiles compared to mutations that compromise the function of the CYP710A protein, as shown in other plant species, the effect of mutations outside of the examples taught by the instant disclosure would be unpredictable to one of ordinary skill in the art. Because undue experimentation would be required to ascertain which modifications would result in reduced activity or expression, and which reductions in activity or expression might result in the claimed sitosterol levels, the instant disclosure does not enable one of ordinary skill in the art to practice the invention over the full scope of the claims.
B. Claims 1-3, 5, 7-8, 10-19, 21-22, 24-32 are drawn to a modified maize plant or progeny or part or cell comprising reduced expression or activity of an endogenous maize CYP710A gene encoding a polypeptide with a sequence with at least 95% sequence identity to SEQ ID NO: 2, or a commodity product produced from such a plant, and a method for producing a maize plant comprising reducing expression or activity of such an endogenous CYP710A gene in the plant. This rejection pertains to making maize plants comprising an endogenous CYP710A gene encoding a polypeptide with as little as 95% sequence identity to SEQ ID NO: 2.
Claims 1-3, 5, 7-8, 10-15, 17-19, 21-22, 24-28 & 31-32 are broadly drawn to a modified maize plant comprising reduced expression or activity of an endogenous CYP710A gene wherein the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 and methods for producing such a plant or producing a commodity plant product from such a plant. Claims 29-30 are drawn to a commodity plant product prepared from the plant, plant part, or plant cell. Claims 2 & 14 require that the CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2 and claims 3 & 15 require that the endogenous CYP710A gene comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO: 1.
An endogenous CYP710A gene encompasses any maize CYP710A gene, including any and all paralogs. Although the claims require reduced expression or activity of an endogenous CYP710A gene and one or more nucleotide modifications at a genomic locus comprising the CYP710A gene, the claims do not require that the nucleotide modifications in the CYP710A gene result in the reduced expression or activity.
A polypeptide comprising an amino acid sequence having at least 95% or 99% sequence identity to the 513 amino acid-long SEQ ID NO: 2 encompasses sequences with 25 or 5 amino acid substitutions relative to SEQ ID NO: 2. Nucleotide sequences with 95% identity to the 1542 nucleotide long SEQ ID NO:1 would have 77 nucleotide substitutions relative to SEQ ID NO: 1.
The instant specification teaches an example of a maize plant with a mutation in a CYP710A gene having increased sitosterol (paragraph [0260]). The instant specification teaches that maize has a single gene model homologous to AtCYP710A1 and responsible for stigmasterol biosynthesis in maize (paragraph [0238 & 0245-0246]). The instant specification teaches that this gene, given the name ZmCYP710A8, is the sole maize C-22 sterol desaturase to synthesize stigmasterol (paragraph [0018], figure 5), and is the Zm00001d039384 gene (paragraph [0238]).
The specification teaches one example of a CYP710A gene in maize as SEQ ID NO:1, which encodes SEQ ID NO:2. The specification also provides the nucleotide sequence SEQ ID NO: 3 which is not described but appears to also encode the CYP710A protein and has 90.4% sequence similarity to SEQ ID NO: 1. See alignment above. The specification provides the names of 23 plant homologs of CYP710A in 13 species (figure 1C, paragraph [0239]), but the instant claims are drawn to an endogenous gene in a maize plant, which excludes homologs of other species.
Endogenous CYP710A genes that are maize CYP710A genes and are known in the art do not have high sequence variability. UniProtKB entry A0A3L6FJ92_MAIZE (available 2/13/2019) describes a CYP710A11 protein in maize with 98.9% sequence identity to the CYP710A8 protein of the instant specification, comprising only one mismatch and 5 indels. See alignment below. Furthermore, the protein of UniProtKB entry A0A3L6FJ92_MAIZE, Zm00014a000506, despite having >95% sequence identity to SEQ ID NO: 2, is not the CYP710A8 gene described by the instant specification, and therefore would not be expected to fulfill the stigmasterol synthesis role of CYP710A8 despite the high sequence similarity according to the teachings of the instant specification.
A0A3L6FJ92_MAIZE
ID A0A3L6FJ92_MAIZE Unreviewed; 518 AA.
AC A0A3L6FJ92;
DT 13-FEB-2019, integrated into UniProtKB/TrEMBL.
DT 13-FEB-2019, sequence version 1.
DT 27-NOV-2024, entry version 23.
DE RecName: Full=C-22 sterol desaturase {ECO:0000256|ARBA:ARBA00041546};
GN Name=CYP710A11 {ECO:0000313|EMBL:PWZ33314.1};
GN ORFNames=Zm00014a_000506 {ECO:0000313|EMBL:PWZ33314.1};
OS Zea mays (Maize).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Liliopsida; Poales; Poaceae; PACMAD clade;
OC Panicoideae; Andropogonodae; Andropogoneae; Tripsacinae; Zea.
OX NCBI_TaxID=4577 {ECO:0000313|EMBL:PWZ33314.1, ECO:0000313|Proteomes:UP000251960};
RN [1] {ECO:0000313|EMBL:PWZ33314.1, ECO:0000313|Proteomes:UP000251960}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Missouri 17 {ECO:0000313|Proteomes:UP000251960};
RC TISSUE=Seedling {ECO:0000313|EMBL:PWZ33314.1};
RX PubMed=30061735; DOI=10.1038/s41588-018-0182-0;
RA Sun S., Zhou Y., Chen J., Shi J., Zhao H., Zhao H., Song W., Zhang M.,
RA Cui Y., Dong X., Liu H., Ma X., Jiao Y., Wang B., Wei X., Stein J.C.,
RA Glaubitz J.C., Lu F., Yu G., Liang C., Fengler K., Li B., Rafalski A.,
RA Schnable P.S., Ware D.H., Buckler E.S., Lai J.;
RT "Extensive intraspecific gene order and gene structural variations between
RT Mo17 and other maize genomes.";
RL Nat. Genet. 50:1289-1295(2018).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5-dehydroepisterol + NADPH + O2 + H(+) =
CC ergosta-5,7,22,24(28)-tetraen-3beta-ol + NADP(+) + 2 H2O;
CC Xref=Rhea:RHEA:33467, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:18249, ChEBI:CHEBI:52972,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.14.19.41;
CC Evidence={ECO:0000256|ARBA:ARBA00036102};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000256|ARBA:ARBA00001971,
CC ECO:0000256|PIRSR:PIRSR602401-1};
CC -!- SIMILARITY: Belongs to the cytochrome P450 family.
CC {ECO:0000256|ARBA:ARBA00010617, ECO:0000256|RuleBase:RU000461}.
CC -!- CAUTION: The sequence shown here is derived from an EMBL/GenBank/DDBJ
CC whole genome shotgun (WGS) entry which is preliminary data.
CC {ECO:0000313|EMBL:PWZ33314.1}.
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DR EMBL; NCVQ01000004; PWZ33314.1; -; Genomic_DNA.
DR AlphaFoldDB; A0A3L6FJ92; -.
DR Proteomes; UP000251960; Chromosome 3.
DR ExpressionAtlas; A0A3L6FJ92; baseline and differential.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR CDD; cd11082; CYP61_CYP710; 1.
DR FunFam; 1.10.630.10:FF:000021; Cytochrome P450 61; 1.
DR Gene3D; 1.10.630.10; Cytochrome P450; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR PANTHER; PTHR24286:SF228; C-22 STEROL DESATURASE ERG5; 1.
DR PANTHER; PTHR24286; CYTOCHROME P450 26; 1.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; Cytochrome P450; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 3: Inferred from homology;
KW Heme {ECO:0000256|PIRSR:PIRSR602401-1, ECO:0000256|RuleBase:RU000461};
KW Iron {ECO:0000256|ARBA:ARBA00023004, ECO:0000256|PIRSR:PIRSR602401-1};
KW Membrane {ECO:0000256|ARBA:ARBA00022989};
KW Metal-binding {ECO:0000256|ARBA:ARBA00022723,
KW ECO:0000256|PIRSR:PIRSR602401-1};
KW Monooxygenase {ECO:0000256|RuleBase:RU000461};
KW Oxidoreductase {ECO:0000256|ARBA:ARBA00023002,
KW ECO:0000256|RuleBase:RU000461};
KW Transmembrane {ECO:0000256|ARBA:ARBA00022692};
KW Transmembrane helix {ECO:0000256|ARBA:ARBA00022989}.
FT BINDING 458
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000256|PIRSR:PIRSR602401-1"
SQ SEQUENCE 518 AA; 57554 MW; ED064B8EFA8FB03E CRC64;
Query Match 98.9%; Score 2665.5; Length 518;
Best Local Similarity 98.8%;
Matches 512; Conservative 0; Mismatches 1; Indels 5; Gaps 2;
Qy 1 MAAQSLS--VPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLG 58
||||||| ||||| |||||||||||||||||||||||||||||||||||||||||||||
Db 1 MAAQSLSVPVPVPVPSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLG 60
Qy 59 SVAHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHL 118
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 SVAHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHL 120
Qy 119 IGHPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDR 178
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 IGHPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDR 180
Qy 179 S---GGGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPC 235
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 SGGGGGGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPC 240
Qy 236 FAFRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPP 295
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 FAFRRAREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPP 300
Qy 296 AHTDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPM 355
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 AHTDDEEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPM 360
Qy 356 TAETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQ 415
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TAETIQGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQ 420
Qy 416 GFPDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVD 475
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 GFPDAEAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVD 480
Qy 476 FRRERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 513
||||||||||||||||||||||||||||||||||||||
Db 481 FRRERTPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 518
Because maize comprising endogenous maize CYP710A genes encoding a polypeptide with 95% or 99% identity to SEQ ID NO: 2 or with 95% sequence identity to SEQ ID NO: 1 are not known in the art or taught by the specification over the full scope of the claimed sequences, one of ordinary skill in the art would be required to conduct undue experimentation to generate maize plants comprising endogenous CYP710A genes across the genus of claimed sequence identities and then to further modify said maize plants to reduce expression or activity of the novel endogenous CYP710A genes in order to practice the invention as claimed. Because the instant specification does not provide examples or guidance on generating maize plants comprising endogenous CYP710A genes over the full scope of the claims, one of ordinary skill in the art would require trial and error experimentation to produce such plants, twice modified, and determine which, if any, would have increased sitosterol or hard endosperm. Claims 1-3, 5, 7-8, 10-19, 21-22 & 24-32 are not enabled over the full scope of any maize plant with an endogenous CYP710A gene encoding a polypeptide with 95% or 99% identity to SEQ ID NO: 2 or with 95% sequence identity to SEQ ID NO: 1.
Applicant urges that the amended claims recite the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, which is more narrowly tailored to the working examples in the specification and encompass only sequences having high sequence similarity to SEQ ID NO: 2 (Remarks, page 7, paragraph 3-page 8, paragraph 1).
This argument is unpersuasive, because the claims require an endogenous maize CYP710A gene encoding a sequence with at least 95% sequence identity to SEQ ID NO: 2, but the instant specification and the prior art do not teach examples of maize plants comprising endogenous sequences over the full scope of the claims. Endogenous maize CYP710A genes encoding a sequence with 25 or even 5 amino acid substitutions are not known in the art or provided by the instant application, so one of ordinary skill in the art would have to discover such CYP710A genes in maize plants, if such endogenous CYP710A genes exist, or create maize plants comprising this broadly claimed genus, if sequences over this full genus can even be considered endogenous maize CYP710A genes. Generating the maize plants comprising the full scope of the claimed endogenous maize CYP710A genes required by the instant claims would require undue trial and error experimentation and is not fully enabled.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claim(s) 1-3, 5, 10-15, 19 & 24-26 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by McCarty et al (2005) The Plant Journal. 44: 52-61 (published 9/6/2005, hereafter McCarty), taken with the evidence of MaizeGDB stock record UFMu-03461 (accessed 6/11/2026), MaizeGDB Jbrowse UniformMu track for region chr3:3906076..3912315 (accessed 6/11/2026), W22 NRGene 2.0 CDS BLAST result (accessed 6/11/2026), and W22 NRGene 2.0 translations BLAST result (accessed 6/11/2026).
Claims 1-3, 5, 10-12 are drawn to a modified maize plant comprising increased sitosterol and reduced expression or activity of an endogenous CYP710A gene wherein the endogenous CYP710A gene encodes a polypeptide comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 2. Claims 13-15, 19 & 24-26 are drawn to methods of producing a modified maize plant.
McCarty discloses the construction of the UniformMu population by introgression of active Mutator and the bz1-mum9 gene into line W22 and backcrossing 8 times to purge parental seed mutants each generation (page 53, right column, paragraph 2; page 59, left column, paragraph 3-4). McCarty discloses that some Mu insertions cause suppressible phenotypes that depend on the presence of Mu activity (page 54, right column, paragraph 2). McCarty discloses a public resource of over 2000 seed mutants to be deposited at the Maize Stock Center and that the method can be applied to other plant processes or lines (page 56, right column, paragraph 3-page 57, left column, paragraph 1). McCarty discloses that the UniformMu population includes mutagenic activity and genetic control of Mu transposition (page 53, left column, paragraph 4).
MaizeGDB stock record UFMu-03461 provides evidence that the UniformMu stock line UFMu-03461 comprises the mu1034077::Mu variation (see Overview).
The MaizeGDB Jbrowse UniformMu track for region chr3:3906076..3912315 provides evidence that the mu1034077 variation is found within a coding sequence of a locus annotated to be a Cytochrome P450 710A1 gene.
W22 NRGene 2.0 CDS BLAST result provides evidence that W22 lines comprise a DNA sequence with 100% sequence identity to instant SEQ ID NO: 1 at target ID Zm00004b015707. See alignment below.
Alignment details for hit #1
Score = 1542 bits (2848), Expect =
Identities = 1542/1542 (1.0000%), Gaps = (0.0000%)
Strand = Minus / Plus
Query 1 ATGGCGGCGCAGTCACTGTCCGTGCCCGTGCCCGTGCTCAGCTCCTTGGACCTCCGCGCG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3868806 ATGGCGGCGCAGTCACTGTCCGTGCCCGTGCCCGTGCTCAGCTCCTTGGACCTCCGCGCG 3868865
Query 61 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCCTTGTACTTCCTGATGGAGCAGGTGTCCTAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3868866 GCGGCGCCGTTCCTGGTTGCCGCGGTGGCCTTGTACTTCCTGATGGAGCAGGTGTCCTAC 3868925
Query 121 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3868926 CACCGGAAGAAGGGGCCCCTGCCCGGGCCGGCGCTCGTGGTGCCGTTCCTCGGCAGCGTG 3868985
Query 181 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3868986 GCGCACATGATCCGCGACCCGACGGGGTTCTGGGACGCGCAGGCGGCGCGCGCCAAGCGG 3869045
Query 241 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869046 TCCGGCGCGGGCCTCGCCGCCGACTTCCTGGTGGGGCGGTTCGTGGTGTTCATCCGCGAC 3869105
Query 301 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869106 TCGGAGCTGTCCCACCGCGTGTTCGCCAACGTCCGCCCCGACGCGTTCCACCTCATCGGC 3869165
Query 361 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869166 CACCCCTTCGGCAAGAAGCTGTTCGGCGACCACAACCTCATCTACATGTTCGGCGAGGAC 3869225
Query 421 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869226 CACAAGGACCTGCGGCGGCGGATCGCGCCCAACTTCACCCCGCGCGCGCTGTCCACCTAC 3869285
Query 481 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869286 GCCGCGCTCCAGCAGCGCGTCATCCTGGCGCACCTCCGGCGGTGGCTCGACCGGAGCGGC 3869345
Query 541 GGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACCTCGCAGACG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869346 GGCGGGGCGTTCCCCATCCGCGTGCCCTGCCGCGACATGAACCTGGAGACCTCGCAGACG 3869405
Query 601 GTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGGGACTACAAC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869406 GTGTTCGTGGGGCCGTACCTCGCCGGGGAGGCCCGGCGGCGGTTCGAGCGGGACTACAAC 3869465
Query 661 CTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCGTTCCGGCGC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869466 CTCTTCAACGTCGGGCTCATGGCGCTGCCCATCGACCTGCCCTGCTTCGCGTTCCGGCGC 3869525
Query 721 GCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGGCAGAGCAAG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869526 GCCAGGGAGGGCGTGGCGCGGCTGGTGCGCACGCTGGGCGAGTGCGCGCGGCAGAGCAAG 3869585
Query 781 GCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATGCAGGACACG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869586 GCGCGGATGCGCGCCGGCGGCGAGCCCGAGTGCCTGGTGGACTTCTGGATGCAGGACACG 3869645
Query 841 CTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCACACCGACGAC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869646 CTGCGGGAGATGGACGAGGCGGCGGCGGCGGGGCGCCCGCCCCCCGCGCACACCGACGAC 3869705
Query 901 GAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCGTCCACCTCG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869706 GAGGAGATCGGCGGGTTCATGTTCGACTTCCTGTTCGCGGCGCAGGACGCGTCCACCTCG 3869765
Query 961 TCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCCCGCGTGCGC 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869766 TCCCTCTGCTGGGCGGTGTCGGCGCTGGACTCCCACCCGGAGGTGCTGGCCCGCGTGCGC 3869825
Query 1021 GCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCGGAGACGATC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869826 GCCGAGGTGTCGGCCGCCTGGTCGCCGGACTCCGGCGAGCCGATGACGGCGGAGACGATC 3869885
Query 1081 CAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGGCCCCCCGCG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869886 CAGGGGATGCGGTACACGCAGGCGGTGGCGCGCGAGGTGGTCCGGCACCGGCCCCCCGCG 3869945
Query 1141 ACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTACACGGTGCCC 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3869946 ACGCTGGTGCCGCACATCGCGGGCGAGGCGTTCCAGCTGACGGAGTGGTACACGGTGCCC 3870005
Query 1201 AAGGGCGCCATCGTGTTCCCGTCCGTGTACGAGTCGTCGTTCCAGGGGTTCCCGGACGCG 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870006 AAGGGCGCCATCGTGTTCCCGTCCGTGTACGAGTCGTCGTTCCAGGGGTTCCCGGACGCG 3870065
Query 1261 GAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTGGCGTACAGG 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870066 GAGGCGTTCGACCCGGAGCGCTTCTTCTCGGAGGCGCGGCGGGAGGACGTGGCGTACAGG 3870125
Query 1321 CGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAGCGGTACGCGCTG 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870126 CGCAACTTCCTGGCATTCGGCGCCGGGCCGCACCAGTGCGTGGGGCAGCGGTACGCGCTG 3870185
Query 1381 AACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTGGACTTCCGGCGGGAGAGG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870186 AACCACCTCGTCCTGTTCATGGCGCTGTTCGTGTCGGTGGTGGACTTCCGGCGGGAGAGG 3870245
Query 1441 ACGCCCGGGTGTGATGACCCGGTGTACATGCCCACCATCGTGCCCAAGGACGGCTGCTCC 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870246 ACGCCCGGGTGTGATGACCCGGTGTACATGCCCACCATCGTGCCCAAGGACGGCTGCTCC 3870305
Query 1501 GTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 1542
||||||||||||||||||||||||||||||||||||||||||
Sbjct 3870306 GTGTACCTCAAGCAGCGATGCGCCAAGTTGCCGTCGTTCTGA 3870347
W22 NRGene 2.0 translations BLAST result provides evidence that target Zm00004b015707_P001 encodes a protein with 100% sequence identity to instant SEQ ID NO: 2. See alignment below.
Alignment details for hit #1
Score = 2704 bits (1046), Expect =
Identities = 513/513 (1.0000%), Gaps = (0.0000%)
Strand = Minus / Plus
Query 1 MAAQSLSVPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
MAAQSLSVPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV
Sbjct 1 MAAQSLSVPVPVLSSLDLRAAAPFLVAAVALYFLMEQVSYHRKKGPLPGPALVVPFLGSV 60
Query 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG
Sbjct 61 AHMIRDPTGFWDAQAARAKRSGAGLAADFLVGRFVVFIRDSELSHRVFANVRPDAFHLIG 120
Query 121HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG 180
HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG
Sbjct 121HPFGKKLFGDHNLIYMFGEDHKDLRRRIAPNFTPRALSTYAALQQRVILAHLRRWLDRSG 180
Query 181GGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFAFRR 240
GGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFAFRR
Sbjct 181GGAFPIRVPCRDMNLETSQTVFVGPYLAGEARRRFERDYNLFNVGLMALPIDLPCFAFRR 240
Query 241AREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAHTDD 300
AREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAHTDD
Sbjct 241AREGVARLVRTLGECARQSKARMRAGGEPECLVDFWMQDTLREMDEAAAAGRPPPAHTDD 300
Query 301EEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTAETI 360
EEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTAETI
Sbjct 301EEIGGFMFDFLFAAQDASTSSLCWAVSALDSHPEVLARVRAEVSAAWSPDSGEPMTAETI 360
Query 361QGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGFPDA 420
QGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGFPDA
Sbjct 361QGMRYTQAVAREVVRHRPPATLVPHIAGEAFQLTEWYTVPKGAIVFPSVYESSFQGFPDA 420
Query 421EAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFRRER 480
EAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFRRER
Sbjct 421EAFDPERFFSEARREDVAYRRNFLAFGAGPHQCVGQRYALNHLVLFMALFVSVVDFRRER 480
Query 481TPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 513
TPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF
Sbjct 481TPGCDDPVYMPTIVPKDGCSVYLKQRCAKLPSF 513
The UniformMu lines of McCarty include a mutation line, UFMu-03461, comprising an insertion mutation at a gene that reads on a nucleotide modification in a coding region of a CYP710A gene. The UniformMu lines, being in a W22 background, read on lines comprising a gene comprising a sequence with 100% sequence identity to SEQ ID NO: 1 encoding a protein with 100% sequence identity to SEQ ID NO: 2. Additionally, because instant SEQ ID NO: 1 has > 80% sequence identity to SEQ ID NO: 3, claim 10 is anticipated.
Although the art is silent with respect to increased sitosterol or hard endosperm of UFMu-03461, maize plants of this line comprising the disclosed mutation in a CYP710A gene would inherently have sitosterol increased, and increased at least 1.2-fold in the seed of the maize plant. Thus, McCarty’s UniformMu lines anticipate instant claims 1-3, 5 & 11. McCarty’s disclosure of deposited seed read on a seed of the modified maize plant of claim 12.
The method of generating UniformMu lines such as UFMu-03461 via transposon mutagenesis reads on a method for producing a modified maize plant comprising reducing expression or activity of an endogenous CYP710A gene of claim 13. The method to modify a maize plant that resulted in line UFMu-03461 comprised the endogenous CYP710A gene comprising a sequence having 100% sequence identity to SEQ ID NO: 1 and encoding a polypeptide having 100% identity to SEQ ID NO: 2 and the modification was introduced within a coding region (instant claims 13-15 & 19). Although the art is silent with respect to increased sitosterol or hard endosperm of UFMu-03461, maize plants of this line comprising the disclosed mutation in a CYP710A gene would inherently have sitosterol increased, and increased at least 1.2-fold in the seed of the maize plant (claim 24). Finally, the backcrossing of the UniformMu lines reads on a method comprising crossing the maize plant with another maize plant to produce seed and growing a progeny maize plant from the seed (claim 25) and crossing the progeny maize plant with itself or another maize plant and repeating the steps of growing and crossing the progeny (claim 26).
Claims 1-3, 5, 10-15, 19 & 24-26 are anticipated by McCarty because the claims do not require any limitations not found in the UniformMu UFMu-03461 line that was available prior to the filing of the instant application and the method of making UFMu-03461.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claim(s) 1-3, 5, 10-15, 19 & 24-28 are rejected under 35 U.S.C. 103 as being unpatentable over McCarty et al (2005) The Plant Journal. 44: 52-61 (published 9/6/2005, hereafter McCarty), taken with the evidence of MaizeGDB stock record UFMu-03461 (accessed 6/11/2026), MaizeGDB Jbrowse UniformMu track for region chr3:3906076..3912315 (accessed 6/11/2026), W22 NRGene 2.0 CDS BLAST result (accessed 6/11/2026), and W22 NRGene 2.0 translations BLAST result (accessed 6/11/2026) in view of McCarty et al (2013) Chapter 11. Thomas Peterson (ed.), Plant Transposable Elements: Methods and Protocols, Methods in Molecular Biology. 1057. 157-166. (available 1/1/2013, hereafter McCarty 2013).
Claims 27-28 are drawn to a maize crop comprising a plurality of maize plants of claim 1 in an agricultural field and a method comprising cultivating a plurality of plants as a maize crop.
The teachings of McCarty, MaizeGDB stock record UFMu-03461, MaizeGDB Jbrowse UniformMu track for region chr3:3906076..3912315, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result are presented above. They do not teach cultivating a plurality of modified maize plants in an agricultural field as a maize crop.
McCarty 2013 teaches that the UniformMu transposon population is a resource that distributes seed stocks carrying germinal insertions (page 157, paragraph 1). McCarty 2013 teaches that the non-transgenic UFMu insertion lines are screened for loss of Mutator activity and deposited in the Maize Genetics Cooperation Stock Center (page 159, paragraph 1). W22 is the recurrent parent for UniformMu lines (page 159, paragraph 2). McCarty 2013 teaches that a fraction of Mu-induced mutations, typically mutations near the 5’-end of genes, have mutant phenotypes only when MuDR is also present in the genome (page 159, paragraph 3). McCarty 2013 teaches that 15 seeds are provided for each request of a UFMu line, which are pooled from ears of between 2 and 5 sib-pollinated F3 plants. McCarty 2013 teaches that sib-pollination minimizes selection against deleterious phenotypes and maximizes recovery of insertions (page 161, paragraph 2).
Before the filing of the instant application, it would have been obvious to one of ordinary skill in the art to plant a plurality of the modified maize plants together in an agricultural field, cultivate as a crop, and harvest seeds following the method of McCarty 2013 to grow sib-pollinated UFMu lines to provide seeds for UniformMu stock requests. One of ordinary skill in the art would have been motivated to combine the method of McCarty 2013 to grow a UniformMu line of McCarty because sib-pollination minimizes selection against deleterious phenotypes and maximizes recovery of insertions. One of ordinary skill in the art would have had reasonable expectation of success, because both methods use UniformMu maize lines.
Although McCarty is silent with respect to the seeds having increased sitosterol or hard endosperm, seed of the UFMu-03461 line would inherently have this phenotype due to the mutation in the CYP710A gene.
Claims 1-3, 5, 10-15, 19 & 24-26 have been mapped to McCarty above. Claims 1-3, 5, 10-15, 19 & 24-28 are obvious in view of McCarty and McCarty 2013.
Claim(s) 1-3, 5, 11-15, 19, & 24 are rejected under 35 U.S.C. 103 as being unpatentable over Griebel et al (2010) The Plant Journal. 63: 254–268 (published 5/18/2010, hereafter Griebel) in view of Gonzalez et al (1979) Current Microbiology. 2: 75-80 (published March, 1979, hereafter Gonzalez), W22 NRGene 2.0 CDS BLAST result (accessed 6/11/2026), and W22 NRGene 2.0 translations BLAST result (accessed 6/11/2026).
Griebel teaches that Arabidopsis genes are strongly increased in P. syringae-inoculated leaves, including the P450 enzyme CYP710A1 responsible for C22 desaturation of β-sitosterol. An increased stigmasterol to β-sitosterol ratio in leaves attenuates specific plant defense responses, which results in enhanced susceptibility to P. syringae (page 255, left column, paragraph 2-right column, paragraph 2).
Arabidopsis lines with T-DNA insertions in the promoter region of the CYP710A1 gene are impaired in P. syringae-induced increase of CYP710A transcript levels and accumulation of stigmasterol, and this reaction is confined to the C24 ethylcholesterol β-sitosterol (page 256, left column, paragraph 2-right column, paragraph 1). Cyp710A mutant plants had lower multiplication of compatible P. syringae in leaves and enhanced resistance with no affect on systemic acquired resistance; a higher ratio of stigmasterol to β-sitosterol negatively affects resistance to P. syringae strains in Arabidopsis (page 257, left column, paragraph 2-right column 2; figure 4).
Griebel teaches that CYP710A-mediated C22 desaturation of β-sitosterol to stigmasterol is evolutionarily conserved among land plants (page 263, left column, paragraph 2).
Griebel does not teach a maize plant comprising a modified endogenous CYP710A gene encoding a polypeptide with 95% sequence identity to SEQ ID NO: 2.
Gonzalez teaches that a strain of Pseudomonas syringae causes holcus spot disease in maize (page 75, left column, paragraph 3; figure 5).
The teachings of W22 NRGene 2.0 CDS BLAST result and W22 NRGene 2.0 translations BLAST result are presented above.
Before the time of filing of the instant application, one of ordinary skill in the art would have been motivated to modify the method of Griebel of using a knockout CYP710A line to reduce susceptibility to P. syringae to maize. One of ordinary skill in the art would have been motivated to try knocking CYP710A in maize to reduce holcus spot disease. One of ordinary skill in the art would have had reasonable expectation of success because CYP710A-mediated C22 desaturation of β-sitosterol to stigmasterol is evolutionarily conserved among land plants.
It would have been obvious to perform the knockout in a common maize cultivar such as W22, which comprises a sequence having 100% sequence identity to SEQ ID NO: 1 and encodes a polypeptide having 100% identity to SEQ ID NO: 2. It would have been obvious to introduce an insertion into a promoter region to knock out the gene, as taught by the cyp710a mutants in Arabidopsis. A lower ratio of stigmasterol to β-sitosterol after infection reads on increased sitosterol. It would have been obvious to search mutants for increased sitosterol, and some of those created by an insertion mutation into CYP710A would have sitosterol increased at least 1.2-fold in the seed of the maize plant (claim 24).
Thus, the modified maize plants and the methods of making them of claims 1-3, 5, 11-15, 19, & 24 are obvious in light of Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result.
Claim(s) 17-18 are rejected under 35 U.S.C. 103 as being unpatentable over Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result as applied to claims 1-3, 5, 11-15, 19, & 24 above, and further in view of Doll et al (2019) Plant Cell Reports. 38: 487-501 (published 1/25/2019, hereafter Doll).
Claims 17-18 are drawn to a method for producing a modified maize plant comprising nucleotide modifications introduced through targeted DNA modification and/or use of a guide RNA and an RNA-guided endonuclease.
The teachings of Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result are presented above. They do not teach targeted DNA modification or use of a guide RNA and an RNA-guided endonuclease.
Doll teaches that maize is a major crop species (page 488, left column, paragraph 1). Doll teaches that although a large number of natural maize mutants exist, increasing diversity through mutagenesis has been a long-term goal (page 488, left column, paragraph 2). Doll teaches that targeted mutagenesis, such as using CRISPR-Cas9 technology, gives ready access to specific mutants; CRISPR-Cas9 technology comprises recognition of DNA due to a short guide RNA that forms a complex with the Cas9 protein (page 488, left column, paragraph 3-right column, paragraph 2). Doll teaches that CRISPR-Cas9 technology is efficient and user-friendly and has been successfully adapted to maize (page 489, left column, paragraphs 1-2).
Prior to the filing of the instant application, one of ordinary skill in the art would have been motivated o modify the method of generating CYP710A knockouts taught by Griebel (T-DNA insertions) for a targeted mutagenesis technology such as CRISPR-Cas9. One of ordinary skill in the art would have been motivated to use CRISPR-Cas9 because it is efficient. One of ordinary skill in the art would have had reasonable expectation of success, because the technology had already been adapted to maize.
In light of Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, W22 NRGene 2.0 translations BLAST result, and Doll, claims 1-3, 5, 11-15, 17-19, & 24 are obvious.
Claim(s) 7-8, 21-22 & 25-26 are rejected under 35 U.S.C. 103 as being unpatentable over Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result as applied to claims 1-3, 5, 11-15, 19, & 24 above, and further in view of McGinnis et al (2007) Plant Physiology. 143: 1441–1451 (published 2/16/2007, hereafter McGinnis).
Claims 7-8 & 21-22 are drawn to a modified maize plant comprising a silencing element that targets the endogenous CYP710A gene or a method wherein the silencing element reduces expression or activity of the CYP710A gene. Claims 25-26 are drawn to methods of producing seeds comprising crossing the maize plant and growing a progeny maize plant from the seed.
The teachings of Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result are presented above. They do not teach a silencing element targeting an endogenous CYP710A gene.
McGinnis teaches a method of using RNA interference (page 1442,let column, paragraph 2, table 1). McGinnis teaches a motivation to use RNAi for loss-of-function, because RNAi-induced mutations are dominant and can induce gene silencing at a transcriptional or posttranscriptional level depending on the sequence in the double-stranded RNA that is processed into siRNA (page 1441, right column, paragraph 2-3). McGinnis teaches a method wherein the transgenic silencing events were outcrossed for multiple generations (page 1443, right column, paragraph 2 & page 1449, left column, paragraph 4). McGinnis teaches that RNAi is a widely used technology (page 1449, left column, paragraph 2).
Before the filing of the instant application, it would have been obvious to generate maize with reduced expression of cyp710A using a silencing element targeting CYP710A such as siRNA. One of ordinary skill in the art would have been motivated to introduce a silencing element into the maize plant, because the RNA interference-generated loss of function would be dominant and could induce gene silencing at a transcriptional or posttranscriptional level. One of ordinary skill in the art would have had reasonable expectation of success, because McGinnis’ siRNA method of generating loss of function mutants was in maize and RNAi was a widely used technology.
Furthermore, growing and crossing the RNAi maize line for multiple generations reads on a method comprising crossing the maize plant to produce seed, growing a plant from the seed, crossing the progeny with another maize plant, and repeating for an additional 0-7 generations (claims 25-26).
Thus, claims 1-3, 5, 7-8, 11-15, 19, 21-22 & 24-26 would have been obvious over Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, W22 NRGene 2.0 translations BLAST result and McGinnis.
Claim(s) 27-32 are rejected under 35 U.S.C. 103 as being unpatentable over Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result as applied to claims 1-3, 5, 11-15, 19, & 24 above, and further in view of Wu et al (US 2012/0017338A1, published 1/19/2012, hereafter Wu).
Claims 27-28 are drawn to a maize crop comprising a plurality of maize plants and a method of producing seeds comprising cultivating a plurality of the plants as a crop and harvesting seed. Claims 31-32 are drawn to a method for producing a commodity plant product. Claims 29-30 are drawn to a commodity plant product prepared from the maize plant, plant part, or plant cell.
The teachings of Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, and W22 NRGene 2.0 translations BLAST result are presented above. They do not teach cultivating a plurality of the plants as a crop and harvesting seed or processing the maize plant to obtain a commodity plant product.
Wu teaches transgenic maize plants, including plants comprising a DNA sequence encoding a polypeptide (SEQ ID NO: 66893) with 95.3% sequence identity to SEQ ID NO: 2. See alignment above.
Wu teaches that seed of the maize plants may be planted, grown and harvested to produce a crop or terminal crop, which is a plant or plant product that is grown and harvested such as leaf, root, shoot, seed, grain, or the like. A terminal crop is grown for uses other than for use as planting seed to produce subsequent generations (paragraph [0024]). Wu also teaches plant by-products made from a plant or plant product by processes, which include crushed corn, or paper made from corn stocks (paragraph [0037]). Wu teaches screening transgenic maize plants in a field to determine phenotypes (paragraph [0070, 0071, 0074]). Wu also teaches plants with increased oil levels (paragraph [0075]).
Prior to the filing of the instant application, it would have been obvious to one of ordinary skill in the art to grow cyp710A mutant maize plants in an agricultural field, as taught by Wu. One of ordinary skill in the art would have been motivated to cultivate the plants in a field in order to screen for beneficial phenotypes or to produce a plant product. One of ordinary skill in the art would have had reasonable expectation of success, because maize was routinely cultivated in fields to produce commodity plant products long before the filing of the instant application.
Thus, a crop comprising a plurality of the maize plants planted together in an agricultural field (claim 27) a method of producing seeds comprising cultivating a plurality of maize plants as a maize crop and harvesting seeds (claim 28) are obvious over Griebel, Gonzalez, W22 NRGene 2.0 CDS BLAST result, W22 NRGene 2.0 translations BLAST result, and Wu. The seeds harvested from the cyp710A mutant plants would inherently have increased sitosterol. Finally, methods of processing the maize plant to obtain a commodity plant product (claim 31), such as those taught by Wu, would have been obvious because maize is grown to produce many products. Crushed maize seeds read on seed meal and leaves read on fodder, and seeds and leaves would comprise DNA of the plant, which reads on a nucleic acid comprising one or more nucleotide modifications at a genomic locus comprising the CYP710A gene, making the commodity plant products of claims 29-30 and 32 obvious.
Conclusion
No claims are allowed.
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/VICTORIA L DELEO/Examiner, Art Unit 1662
/Anne Kubelik/Primary Examiner, Art Unit 1663
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