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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 3/2/2026 has been entered.
Claim Status
Claims 1, 3-4, 6, 8-12, 15, 17, 20, 22 and 26-27 are pending and being examined.
All previous objections and rejections not set forth below have been withdrawn in view of applicant’s amendments to the claims.
Claim Rejections - 35 USC § 103
Claims 1, 6, 8-10, 15, 17, and 26-27 are rejected under 35 U.S.C. 103 as being unpatentable over Bonning et al. (US 20190239513 A1) in view of Zhao et al. (Implication of whitefly vesicle associated membrane protein-associated protein B in the transmission of Tomato yellow leaf curl virus, 2019, Virology 535:210–217). Due to applicant’s amendments, the rejection is modified from the rejection set forth on pages 8-19 in the Office action dated 05/30/2025.
Claim 1 is drawn to a fusion protein comprising a carrier protein attached to an insect-specific toxin via a peptide linker, wherein the carrier protein is a Tomato yellow leaf curl virus coat protein (TYLCV-CP).
Bonning et al. describes a fusion protein comprising a protease-resistant linker (page 13, para 0121) (as recited in claim 5) fused with Junonia coenia densovirus (JcDNV) coat protein (JcDNV CP) and an insect-specific toxin (page 13, para 0123) including insect-specific neurotoxin (page 13, para 0118, line 19-20) (as recited in claim 8) for controlling its target insect pests, fall armyworm (Spodoptera frugiperda) (abstract). It also describes that the protease-resistant linker ensures that the fusion protein is transported into hemocoel as an intact fusion protein comprising the insect specific toxin (page 13, para 0123). Bonning et al. teaches that for intrahemocoelic proteins such as the coat protein of an insect vectored plant virus can be used to deliver toxins into the hemocoel (page 11, para 0111). It describes a method to control insect pest by expressing toxic proteins with insecticidal activity comprising Hv1a protein (as recited in claim 10) fused with JcDNV CP using a protease resistant linker comprising SEQ ID NO: 4 (page 13, para 119).
However, Bonning et al. does not teach a TYLCV CP.
Zhao et al. describes a fusion protein comprising a TYLCV CP fused with GST protein via a peptide linker (page 212, left column, para 2), as discussed on pages 5-6 in the previous Office action dated 5/30/2025. Zhao et al. also describes that TYLCV CP is the only structural protein known to be involved in the virus transmission (page 1, right column, para 2). Once acquired orally by its insect vector, whitefly (Bemisia tabaci) (as recited in claim 27), TYLCV CP follows a sequential path of stylet-midgut-haemolymph-salivary gland, and in this journey inside whitefly, virus has to cross two major physiological barriers, namely midgut and salivary gland, to be transmitted successfully by the insect vector (page 1, left column, para 1; Fig. 5), implying that TYLCV CP has the ability to pass across the gut epithelium and moves into the hemocoel of the target insect, as recited in claim 26.
Before the effective filing date, it would have been obvious to an ordinarily skilled artisan to modify the method as described by Bonning et al. by replacing JcDNV CP in the fusion protein wherein the fusion protein also comprises an insect-specific neurotoxin, as taught by Bonning et al., with the TYLCV CP, as taught by Zhao et al., with a realistic goal to control target insect pest (whitefly). TYLCV CP is shown to be capable to reach the gut (and hemocoel in the gut) of its target insect pest, whiteflies; just like the JcDNV CP which is capable to reach the gut of its target insect pest, fall army worm.
Before the effective filing date, an ordinarily skilled artisan would have been motivated to express a fusion protein comprising TYLCV CP fused with an insect-specific neurotoxin via an protease resistant linker in a plant with a realistic goal to control target insect pests by enabling the fusion protein comprising the TYLCV CP and the insect-specific toxin protein to pass across the gut epithelium and moves into the hemocoel of the target insect pest.
Regarding claim 6, Bonning et al. describes a protease-resistant linker consisting of SEQ ID NO: 4 (page 13, para 119). Translating the nucleotide sequence of instant SEQ ID NO: 3 would give rise to the following sequence: GDDAPPSPGPDPGPQPPPPPPPSPTPVG. Sequence alignment of the said polypeptide sequence and SEQ ID NO: 4 of Bonning et al. shows 100% identity, as shown below.
Title: AASEQ1_04302025_130720
Perfect score: 173
Sequence: 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
Database : AASEQ2_04302025_130720.pep:*
RESULT 1
AASEQ2_04302025_130720
Query Match 100.0%; Score 173; Length 32;Indels 0; Gaps 0;
Best Local Similarity 100.0%; Matches 28; Conservative 0; Mismatches 0;
Qy 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
||||||||||||||||||||||||||||
Db 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
Thus, SEQ ID NO: 4 of Bonning et al. is encoded by instant SEQ ID NO: 3, and they are functional equivalents.
Regarding instant claim 9, Bonning et al. describes an arthropod-derived neuropeptide, enzyme or hormone (page 13, para 0120, line 9-10).
Regarding claims 15 and 17, Bonning et al. describes several vectors (page 24, para 0227) comprising an expression cassette consisting of various promoters comprising plant expressible constitutive promoters including cauliflower mosaic virus (CaMV), NOS promoter and ubiquitin promoter (page 22, para 0215, line 10-12).
Claims 3-4 are rejected under 35 U.S.C. 103 as being unpatentable over Bonning et al. in view of Zhou et al. as applied to claims 1, 6, 8-10, 15, 17 and 26-27 above, and further in view of Fluentes et al. (Field Trial and Molecular Characterization of RNAi-Transgenic Tomato Plants That Exhibit Resistance to Tomato Yellow Leaf Curl Geminivirus, 2016, MPMI, 29:197–209) .
Claims 3-4 depend from claim 1. Claim 3 is drawn to the fusion protein of claim 1, wherein the coat protein comprises the amino acid sequence set forth in SEQ ID NO: 2. Claim 4 is drawn to coat protein encoded by a nucleotide sequence set forth in SEQ ID NO: 1 encoding the said coat protein.
Bonning et al. in view of Zhao et al. describes a fusion protein comprising a TYLCV CP, a protease resistant linker, and an insect specific toxin, as described above.
However, Bonning et al. in view of Zhao et al. do not describe instant SEQ ID NOs: 1-2.
Fuentes et al. describes a nucleotide sequence comprising 100% sequence identity to instant SEQ ID NO: 1 and encoding a TYLCV CP consisting of SEQ ID NO: 2, as shown below. The TYLCV CP polypeptide sequence is highlighted in grey.
RESULT 1
KM926626
LOCUS KM926626 2781 bp DNA linear VRL 21-FEB-2018
DEFINITION Tomato yellow leaf curl virus isolate Cuba C2+, complete genome.
ACCESSION KM926626
VERSION KM926626.1
SOURCE Tomato yellow leaf curl virus (TYLCV)
ORGANISM Tomato yellow leaf curl virus
Viruses; Monodnaviria; Shotokuvirae; Cressdnaviricota;
Repensiviricetes; Geplafuvirales; Geminiviridae; Begomovirus.
REFERENCE 1 (bases 1 to 2781)
AUTHORS Fuentes,A., Carlos,N., Ruiz,Y., Callard,D., Sanchez,Y.,
Ochagavia,M.E., Seguin,J., Malpica-Lopez,N., Hohn,T., Lecca,M.R.,
Perez,R., Doreste,V., Rehrauer,H., Farinelli,L., Pujol,M. and
Pooggin,M.M.
TITLE Field Trial and Molecular Characterization of RNAi-Transgenic
Tomato Plants That Exhibit Resistance to Tomato Yellow Leaf Curl
Geminivirus
JOURNAL Mol. Plant Microbe Interact. 29 (3), 197-209 (2016)
PUBMED 26713353
REFERENCE 2 (bases 1 to 2781)
AUTHORS Fuentes,A. and Pooggin,M.M.
TITLE Direct Submission
JOURNAL Submitted (14-OCT-2014) Department of Environmental Sciences,
Botany, University of Basel, Hebelstrasse 1, Basel, BaselStadt
4056, Switzerland
FEATURES Location/Qualifiers
source 1..2781
/organism="Tomato yellow leaf curl virus"
/mol_type="genomic DNA"
/isolate="Cuba C2+"
/isolation_source="symptomatic young leaf of the tomato
plant"
/host="Solanum lycopersicum cv. Campbell-28"
/db_xref="taxon:10832"
/geo_loc_name="Cuba: open field near Havana"
/collection_date="20-Sep-2011"
gene 148..498
/gene="V2"
CDS 148..498
/gene="V2"
/codon_start=1
/product="pre-coat protein"
/protein_id="AJG06153.1"
/translation="MWDPLLNEFPESVHGFRCMLAIKYLQSVEETYEPNTLGHDLIRD
LISVVRARDYVEATRRYNHFHARLEGSPKAELRQPIQQPCCCPHCPRHKQATIMDVQA
HVPKAQNIQNVSKP"
gene 308..1084
/gene="V1"
CDS 308..1084
/gene="V1"
/codon_start=1
/product="coat protein"
/protein_id="AJG06154.1"
/translation="MSKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRS
WTYRPMYRKPRIYRMYRSPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGI
THRVGKRFCVKSIYFLGKVWMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNM
FDNEPSTATVKNDLRDRFQVMRKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEA
AKYENHTENALLLYMACTHASNPVYATMKIRIYFYDSISN"
gene complement(1081..1485)
/gene="C3"
CDS complement(1081..1485)
/gene="C3"
/note="REn"
/codon_start=1
/product="replication enhancer"
/protein_id="AJG06155.1"
/translation="MDSRTGELITAPQAENGVFIWEIINPLYFKITDHSQRPFLMNHD
IISIQIRFNHNIRKVMGIHKCFLNFRIWTTLQPQTGRFLRVFRYQVLKYLDSLGVISI
NNVIRAVDHVLYDVLENTINVTEAHDIKYKFY"
gene complement(1226..1633)
/gene="C2"
CDS complement(1226..1633)
/gene="C2"
/codon_start=1
/product="transcriptional transactivator"
/protein_id="AJG06156.1"
/translation="MQPSSPSTSHCSQVSIKVQHKIAKKKPIRRKRVDLDCGCSYYLH
LNCNNHGFTHRGTHHCSSGREWRFYLGDNQSPLFQDNRPQPEAISNEPRHHFHSDKIQ
PQYQEGNGDSQMFSQLPNLDDITASDWSFLKSI"
gene complement(1542..2615)
/gene="C1"
CDS complement(1542..2615)
/gene="C1"
/codon_start=1
/product="replication initiator protein"
/protein_id="AJG06157.1"
/translation="MPRLFKIYAKNYFLTYPNCSLSKEEALSQLQNLETPTNKKYIKV
CREFHENGEPHLHVLIQFEGKYQCKNQRFFDLVSPTRSAHFHPNIQAAKSSTDVKTYV
EKDGDYIDFGVFQIDGRSARGGQQSANDAYAEALNSGSKSEALNILKEKAPKDYILQF
HNLSSNLDRIFSPPLEVYVSPFLSSSFNQVPDELEEWVAENVVSSAARPWRPRSIVIE
GDSRTGKTMWARSLGPHNYLCGHLDLSPKVYSNDAWYNVIDDVDPHYLKHFKEFMGAQ
RDWQSNTKYGKPIQIKGGIPTIFLCNPGPTSSYREYLDEEKNISLKNWALKNATFVTL
YEPLFASINQGPTQDSQEETNKA"
gene complement(2171..2464)
/gene="C4"
CDS complement(2171..2464)
/gene="C4"
/codon_start=1
/product="C4"
/protein_id="AJG06158.1"
/translation="MGSLISMCLSSSRENTSARTNGSSIWYPQPGQHISIQTFRQLRA
QQMSRPTWRKTETTLILEFSKSMADQLEEVSNLPTTHMPKHSIQAVNPRPSIY
Best Local Similarity 100.0%; Query Match 100.0%; Score 771; Length 2781;
Matches 771; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 TCGAAGCGACCAGGCGATATAATCATTTCCACGCCCGTCTCGAAGGTTCGCCGAAGGCTG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 311 TCGAAGCGACCAGGCGATATAATCATTTCCACGCCCGTCTCGAAGGTTCGCCGAAGGCTG 370
Qy 61 AACTTCGACAGCCCATACAGCAACCGTGCTGCTGTCCCCATTGTCCAAGGCACAAACAAG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 371 AACTTCGACAGCCCATACAGCAACCGTGCTGCTGTCCCCATTGTCCAAGGCACAAACAAG 430
Qy 121 CGACGATCATGGACGTACAGGCCCATGTACCGAAAGCCCAGAATATACAGAATGTATCGA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 431 CGACGATCATGGACGTACAGGCCCATGTACCGAAAGCCCAGAATATACAGAATGTATCGA 490
Qy 181 AGCCCTGATGTTCCCCGTGGATGTGAAGGCCCATGTAAAGTCCAGTCTTATGAGCAGCGG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 491 AGCCCTGATGTTCCCCGTGGATGTGAAGGCCCATGTAAAGTCCAGTCTTATGAGCAGCGG 550
Qy 241 GATGATATTAAGCACACTGGTGTTGTTCGTTGTGTTAGTGATGTTACTCGTGGATCTGGA 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 551 GATGATATTAAGCACACTGGTGTTGTTCGTTGTGTTAGTGATGTTACTCGTGGATCTGGA 610
Qy 301 ATTACTCACAGAGTCGGTAAGAGGTTCTGTGTTAAATCGATATATTTTTTAGGTAAAGTC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 611 ATTACTCACAGAGTCGGTAAGAGGTTCTGTGTTAAATCGATATATTTTTTAGGTAAAGTC 670
Qy 361 TGGATGGATGAAAATATCAAGAAGCAGAATCACACTAATCAGGTCATGTTCTTTTTGGTC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 671 TGGATGGATGAAAATATCAAGAAGCAGAATCACACTAATCAGGTCATGTTCTTTTTGGTC 730
Qy 421 CGTGATAGAAGGCCCTATGGAAGCAGTCCAATGGATTTTGGACAGGTTTTTAATATGTTC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 731 CGTGATAGAAGGCCCTATGGAAGCAGTCCAATGGATTTTGGACAGGTTTTTAATATGTTC 790
Qy 481 GATAATGAGCCCAGTACCGCAACTGTGAAGAATGATTTGCGTGATAGGTTTCAAGTGATG 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 791 GATAATGAGCCCAGTACCGCAACTGTGAAGAATGATTTGCGTGATAGGTTTCAAGTGATG 850
Qy 541 AGGAAATTTCATGCTACAGTTATTGGTGGGCCCTCTGGAATGAAGGAACAGGCATTAGTT 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 851 AGGAAATTTCATGCTACAGTTATTGGTGGGCCCTCTGGAATGAAGGAACAGGCATTAGTT 910
Qy 601 AAGAGATTTTTTAAAATTAACAGTCATGTAACTTATAATCATCAGGAGGCAGCCAAGTAC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 911 AAGAGATTTTTTAAAATTAACAGTCATGTAACTTATAATCATCAGGAGGCAGCCAAGTAC 970
Qy 661 GAGAACCATACTGAAAACGCCTTGTTATTGTATATGGCATGTACGCATGCCTCTAATCCA 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 971 GAGAACCATACTGAAAACGCCTTGTTATTGTATATGGCATGTACGCATGCCTCTAATCCA 1030
Qy 721 GTGTATGCAACTATGAAAATACGCATCTATTTCTATGATTCAATATCAAAT 771
|||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1031 GTGTATGCAACTATGAAAATACGCATCTATTTCTATGATTCAATATCAAAT 1081
There is also a 100% sequence identity between instant SEQ ID NO: 2 and the TYLCV CP as taught by Fuentes et al., as shown below.
Title: US-18-701-456-2
Perfect score: 1378
Sequence: 1 SKRPGDIIISTPVSKVRRRL..........SNPVYATMKIRIYFYDSISN 257
Searched: 1 seqs, 258 residues
Database : AASEQ2_04292025_112735.pep:*
RESULT 1
AASEQ2_04292025_112735
Query Match 100.0%; Score 1378; DB 1; Length 258; Mismatches 0;
Best Local Similarity 100.0%; Matches 257; Conservative 0; Indels 0; Gaps 0;
Qy 1 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 61
Qy 61 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 62 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 121
Qy 121 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 122 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 181
Qy 181 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 182 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 241
Qy 241 VYATMKIRIYFYDSISN 257
|||||||||||||||||
Db 242 VYATMKIRIYFYDSISN 258
Before the effective filing date, it would have been obvious to an ordinarily skilled artisan to replace nucleotide sequence encoding JcDNV CP in the fusion protein, as taught by Bonning et al., with the TYLCV CP nucleotide sequence, as taught by Fuentes et al., with a realistic goal to control specific target insect pest, whitefly. The fusion protein comprising the TYLCV CP passes across the gut epithelium and moves into the hemocoel of whitefly, as taught by Zhao et al. Using any specific TYLCV CP is within the experimental design choice of the an ordinarily skilled artisan.
Before the effective filing date, an ordinarily skilled artisan would have been motivated to fuse the TYCLV CP with an insect specific toxin using a peptide linker with a realistic goal to control the target insect pest, whitefly, by enabling the fusion protein comprising the TYLCV CP and the insect-specific toxin protein to pass across the gut epithelium and moves into the hemocoel of whitefly.
Claims 11-12, 20 and 22 are rejected under 35 U.S.C. 103 as being unpatentable over Bonning et al. in view of Zhou et al. as applied to claims 1, 6, 8-10, 15, 17 and 26-27 above, in further view of Fuentes et al., as applied to reject claims 3-4 under 35 U.S.C. 103 above, and in evidence of Liao et al. (Removal of N-terminal methionine from recombinant proteins by engineered E. coli methionine aminopeptidase, 2004, Protein Sci., 13:1802–1810).
Claims 11-12 depends from claim 1 and are drawn to SEQ ID NO: 11-12. SEQ ID NO: 11-12, as recited in claims 11-12, comprise the TYLCV CP sequence fused with the Hv1a insect specific toxin via a protease resistant linker.
Bonning et al. in view of Zhao et al., and in further view of Flentes et al. describes a fusion protein comprising TYLCV CP, wherein the TYLCV CP comprises the instant SEQ ID NO: 2, fused with an insect-specific toxin via a protease resistant peptide linker that can pass across the gut epithelium and moves into the hemocoel of the insect, as described above. The fusion protein comprises protease resistant linker consisting of SEQ ID NO: 4 of Bonning et al., as discussed above. Bonning et al. also describes an insecticidal peptide, Hvla, consisting of SEQ ID NO: 6 (page 11, para 0107).
An amino acid sequence encoding the fusion protein comprising of the TYLCV CP (as described by Fuentes et al.), a protease-resistant linker peptide having the sequence “GDDAPPSPGPDPGPQPPPPPPPSPTPVG” (SEQ ID NO: 4 of Bonning et al.), and the Hv1a insecticidal protein (SEQ ID NO: 6 of Bionning et al.), as described above, excluding the start methionine (M) residues for the fused polypeptides, would give 100% sequence identity to SEQ ID NO: 12, as shown below.
Title: US-18-701-456-12
Perfect score: 1773
Sequence: 1 SKRPGDIIISTPVSKVRRRL..........CSQSCTFKENENGNTVKRCD 322
Searched: 1 seqs, 322 residues
Total number of hits satisfying chosen parameters: 1
RESULT 1
AASEQ2_04292025_153807
Query Match 100.0%; Score 1773; Length 322; Conservative 0; Gaps 0;
Best Local Similarity 100.0%; Matches 322; Mismatches 0; Indel 0
Qy 1 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 60
Qy 61 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 120
Qy 121 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 180
Qy 181 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 240
Linker-polypeptide Hv1a
Qy 241 VYATMKIRIYFYDSISNGDDAPPSPGPDPGPQPPPPPPPSPTPVGSPTCIPSGQPCPYNE 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 VYATMKIRIYFYDSISNGDDAPPSPGPDPGPQPPPPPPPSPTPVGSPTCIPSGQPCPYNE 300
Hv1a
Qy 301 NCCSQSCTFKENENGNTVKRCD 322
||||||||||||||||||||||
Db 301 NCCSQSCTFKENENGNTVKRCD 322
The non-highlighted sequence is the TYLCV CP, the first lighter grey highlighted sequence is the protease resistant linker polypeptide, and the darker grey highlighted sequence is the Hv1a insect specific toxin. It is a standard practice in the art that the translation start or initiator methionine (M) residues are usually not included in making fusion proteins. In fact, removal of the translation initiator N-formyl-methionine or methionine from a fusion or a recombinant protein is often critical for its function and stability (Liao et al.; abstract).
Before the effective filing date, it would have been obvious to an ordinarily skilled artisan to fuse a specific TYLCV CP (as described by Fuentes et al.) with an insect-specific toxin via a protease-resistant linker (as described by Bonning et al.) with a realistic goal to control target insect pests (whiteflies), as described above.
Before the effective filing date of the invention, an ordinarily skilled artisan would have been motivated to express a fusion protein consisting of SEQ ID NO: 12 encoded by the polynucleotide comprising SEQ ID NO: 11, with the realistic goal to control target insect pest (whitefly) population. Using any specific polynucleotide including SEQ ID NO: 11 (as recited in claim 20) would be considered functional equivalent so long the polynucleotide encodes the polypeptide having 100% sequence identity to instant SEQ ID NO: 12, as recited in claim 12.
Regarding claim 22, it would have been obvious to an ordinarily skilled artisan and the artisan would have been motived to introduce SEQ ID NO: 11 in a plant cell and produce transgenic plants a with a realistic goal to express the fusion protein comprising SEQ ID NO: 12 to control target insect pests feeding on the plant or parts thereof.
Double Patenting
Claims 1, 6, 8-10, 15, 17 and 26-27 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 4, 6-7, and 12 of U.S. Patent No. US11140902B2 (hereafter referred to as ‘902B) in view of Bonning et al. (US 2019/0239513 A1) and Zhao et al. Due to applicant’s amendments, the rejection is modified from the rejection set forth on pages 19-29 in the Office action dated 05/30/2025.
Instant claim 1 is drawn to a fusion protein comprising a career protein, which is a tomato yellow leaf curl virus coat protein (TYLCV CP), fused to an insect specific toxin via a peptide linker.
Claim 1 of ’902B recites a fusion protein comprising a carrier protein attached to a second peptide via a peptide linker, wherein the carrier protein is JcDNV CP. Claim 7 of ’902B recites an “insect toxin” (which reads on to “insect specific toxin”) Hv1a (line 3).
However, claims 1 or claim 7 of ‘902B does not teach TYLCV CP as the carrier protein.
Zhao et al. describes a fusion protein comprising TYLCV CP fused with a second protein (GST). It also describes that once acquired orally by whitefly, TYLCV CP follows a sequential path of stylet-midgut-haemolymph (the fluid inside hemocoel)-salivary gland. In this journey inside whitefly, virus coat protein crosses two major physiological barriers, namely midgut and salivary gland, to be transmitted successfully by the vector (page 1, left column, para 1; Fig. 5). Zhao et al. also describes that TYLCV CP is the only structural protein known to be involved in the virus transmission (page 1, right column, para 2) implying that TYLCV CP can pass across the gut epithelium and moves into the hemocoel and haemolymph of the insect.
Bonning et al. describes a fusion protein comprising JcDNV CP attached to a peptide linker and is used to deliver a fused insect specific toxin across the gut epithelium of a target insect pest, fall armyworm. The fusion protein is used for controlling the target insect pest (Abstract).
Before the effective filing date, it would have been obvious to an ordinarily skilled artesian to modify the fusion protein as described in claims 1 of ’902B to replace JcDNV CP with TYLCV CP and fuse the TYCLV CP (as described by Zhao et al.) with an insect-specific toxin Hv1a (as recited in claim 7 or ‘902B) via a protease resistant linker (as taught by Bonning et al.) with a realistic objective to deliver the fusion protein into the insect hemocoel, and, thus, controlling the specific target insect population.
Before the effective filing date, an ordinarily skilled artesian would have been, motivated to fuse a TYLCV CP with an insect-specific toxin Hv1a polypeptide via a peptide linker to deliver the fusion protein into insect hemocoel with a realistic goal to control the target insect pest population.
Regarding instant claim 6, claim 7 of ’092B describes a protease-resistant linker consisting of SEQ ID NO: 4. Translating the nucleotide sequence of instant SEQ ID NO: 3 would give rise to the following sequence: GDDAPPSPGPDPGPQPPPPPPPSPTPVG. Sequence alignment of the said amino acid sequence and SEQ ID NO: 4 of ‘092B shows 100% identity, as shown below.
Title: AASEQ1_04302025_130720
Perfect score: 173
Sequence: 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
Database : AASEQ2_04302025_130720.pep:*
RESULT 1
AASEQ2_04302025_130720
Query Match 100.0%; Score 173; DB 1; Length 32; Indels 0;
Best Local Similarity 100.0%; Matches 28; Conservative 0; Mismatches 0; Gaps 0;
Qy 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
||||||||||||||||||||||||||||
Db 1 GDDAPPSPGPDPGPQPPPPPPPSPTPVG 28
Amino acid sequence consisting of SEQ ID NO: 3 in instant claim 6 is encoded by the nucleotide sequence consisting of SEQ ID NO: 4 of claim 7 of ‘902B and, thus, are functional equivalents.
Regarding instant claims 8-10, claims 6 and 7 of ‘902B recite an insect-specific toxin, Hv1a. Bonning et al. teaches an insect specific neurotoxin (page 6, para 0068, line 11-13; page 13, para 0123, last line). It also teaches that Hv1a is an arthropod-derived neuropeptide (page 11, para 0107, line 1-4).
Regarding instant claims 15 and 17, Bonning et al. describes a vector comprising an expression cassette consisting of plant expressible constitutive promoters including cauliflower mosaic virus (CaMV) and ubiquitin promoters (page 22, para 0215) to express the specific proteins in a plant. Using plant expressible promoters including cauliflower mosaic virus (CaMV) and ubiquitin promoters to express specific proteins in a plant is a widely used, known, and routine method to express any protein(s) including fusion protein(s) in a plant.
Regarding instant claims 26-27, Claim 12 of ‘902B are drawn to a method of controlling target insect pests by feeding the target insect pest fall army worm, Spodoptera frugiperda, with a food source comprising a fusion protein comprising a viral (JcDNV) coat protein attached to an insect specific toxin via a peptide linker, thereby allowing the fusion protein to pass across the gut epithelium of the target insect and act within the hemocoel of the insect after the fusion protein is ingested by the insect. Zhao et al. describes that once acquired orally by whitefly (as recited in claim 27), TYLCV CP enables a fusion protein to pass across the gut epithelium and moves into the hemocoel of the insect. The insecticidal toxin fused to the TYLCV CP would act within the insect hemocoel. It would have been obvious to an ordinarily skilled artisan to develop a method of controlling target insect pests by feeding an insect with a food source comprising a fusion protein comprising TYLCV CP (as taught by Zhou et al.) and insect specific toxin protein fused together via a peptide linker (as taught by claim 12 of ‘092B), wherein upon ingestion, the fusion protein passes across the gut epithelium and moves into the hemocoel of the target insect pest (whitefly), as recited in instant claim 26.
Claims 3-4, 11-12, 20 and 22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 4, 6-7, and 12 of U.S. Patent No. US11140902B2 in view of Bonning et al. and Zhou et al., as applied to reject claims 1, 5-6, 8-10, 15, 17, and 26-27 above under nonstatutory double patenting, and in further view of Fuentes et al.
Instant claim 3 recites a fusion protein comprising the TYLCV CP having the amino acid sequence set forth in instant SEQ ID NO: 2. Instant claim 4 recites a fusion protein comprising the TYLCV CP encoded by the nucleotide sequence set forth in SEQ ID NO: 1.
Claims 1, 4, 6-7, and 12 of U.S. Patent No. US11140902B2 in view of Bonning et al. and Zhou et al., describe a fusion protein comprising the TYLCV CP and an insect-specific toxin protein fused together via a peptide linker to deliver the insecticidal protein into insect hemocoel with a realistic goal to control the target insect pest population, as discussed above.
However, claims 1, 4, 6-7, and 12 of U.S. Patent No. US11140902B2 in view of Bonning et al. and Zhou et al., do not describe a TYLCV CP comprising the instant SEQ ID NO: 2 or instant SEQ ID NO: 1.
Fuentes et al. describes a TYLCV CP comprising 100% sequence identity to instant SEQ ID NO: 2. It also teaches a polynucleotide sequence having 100% sequence identity to instant SEQ ID NO: 1, as discussed above.
Before the effective filing date of the invention, it would have been obvious to an ordinarily skilled artisan to replace the nucleotide sequence encoding the JcDNV CP in the fusion protein, as taught by claim 1 of ‘902B, with the TYLCV CP nucleotide sequence having 100% sequence identity with instant SEQ ID NO: 1 encoding a protein having 100% sequence identity with instant SEQ ID NO: 2, as taught by Fuentes et al., with the realistic goal to control specific target insect pest (i.e., whitefly) by enabling the fusion protein comprising the TYLCV CP and the insect-specific toxin protein, as taught by Bonning et al., to pass across the gut epithelium and moves into the hemocoel of whitefly, as taught by Zhao et al. It is noted that using a functional equivalent of the nucleotide sequence encoding TYLCV CP is within the experimental design choice of the Applicant.
Before the effective filing date, an ordinarily skilled artisan would have been motivated to fuse the TYCLV CP with an insect specific toxin using a peptide linker with the realistic goal to control the target insect pest, whitefly, by enabling the fusion protein comprising the TYLCV CP and the insect-specific toxin protein to pass across the gut epithelium and moves into the hemocoel of whitefly.
Regarding claims 11-12, instant claim 11 is drawn to fusion protein comprising the nucleotide sequence set forth in instant SEQ ID NO: 11 while the instant claim 12 recites the fusion protein of instant claim 1, comprising the amino acid sequence set forth in instant SEQ ID NO: 12. A fusion protein comprising the TYLCV CP, as described by Fuentes et al., the linker peptide having the sequence of SEQ ID NO: 4 as recited in claim 7 of ‘902B, and the Hv1a insect specific toxin having the sequence of SEQ ID NO: 6 of ‘902B without the start methionine (M) would give rise to a fusion protein having 100% sequence identity with SEQ ID NO: 12, as shown below.
Title: US-18-701-456-12
Perfect score: 1773
Sequence: 1 SKRPGDIIISTPVSKVRRRL..........CSQSCTFKENENGNTVKRCD 322
Searched: 1 seqs, 322 residues
Total number of hits satisfying chosen parameters: 1
RESULT 1
AASEQ2_04292025_153807
Query Match 100.0%; Conservative 0; Best Local Similarity 100.0%; Indel 0;
Matches 322; Mismatches 0; Score 1773; Gaps 0; Length 322
Qy 1 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 SKRPGDIIISTPVSKVRRRLNFDSPYSNRAAVPIVQGTNKRRSWTYRPMYRKPRIYRMYR 60
Qy 61 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 SPDVPRGCEGPCKVQSYEQRDDIKHTGVVRCVSDVTRGSGITHRVGKRFCVKSIYFLGKV 120
Qy 121 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 WMDENIKKQNHTNQVMFFLVRDRRPYGSSPMDFGQVFNMFDNEPSTATVKNDLRDRFQVM 180
Qy 181 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 RKFHATVIGGPSGMKEQALVKRFFKINSHVTYNHQEAAKYENHTENALLLYMACTHASNP 240
Linker-polypeptide Hv1a
Qy 241 VYATMKIRIYFYDSISNGDDAPPSPGPDPGPQPPPPPPPSPTPVGSPTCIPSGQPCPYNE 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 VYATMKIRIYFYDSISNGDDAPPSPGPDPGPQPPPPPPPSPTPVGSPTCIPSGQPCPYNE 300
Hv1a
Qy 301 NCCSQSCTFKENENGNTVKRCD 322
||||||||||||||||||||||
Db 301 NCCSQSCTFKENENGNTVKRCD 322
The non-highlighted part is the TYLCV CP, the first lighter grey highlighted sequence is the linker polypeptide, and the darker grey highlighted sequence is the Hv1a insecticidal protein. Translation start or initiator methionine (M) residues are usually not included in fusion proteins. In fact, removal of the translation initiator N-formyl-methionine or methionine from a fusion or a recombinant protein is often critical for its function and stability1.
Any nucleotide sequence (e.g., SEQ ID NO: 11) encoding the said fusion protein comprising SEQ ID NO: 12 would be a functional equivalent and within the experimental design choice of an ordinarily skilled artisan without having any realistic possibility to change the outcome of controlling the target insect pest, whitefly.
Regarding instant claims 20 and 22, It would have been obvious to an ordinarily skilled artisan and the artisan would have been motivated to develop a transgenic plant using any routine and standard technique (e.g. agrobacterium media transformation) and express the nucleotide sequence comprising SEQ ID NO: 11 or its functional equivalent encoding a fusion protein comprising the recombinant polypeptide sequence of instant SEQ ID NO: 12, with a realistic goal to control target insect pest (whitefly) that would be feeding on the transgenic plant or parts thereof, as discussed above.
Response to Applicant’s Arguments
The response from the Applicant dated 3/2/2026 is fully considered but not found persuasive.
In regard to 103 rejection, the Applicant argues, “JcDNV and TYLCV belong to completely different virus families, which have distinct structural characteristics, capsid architectures, and mechanisms of action. A person of ordinary skill in the art would not have been motivated to substitute one coat protein for the other based on these known distinct characteristics” (response, page 5, para 3, line 1-4) and “Zhou reports the movement of the whole virus (and NOT the movement of individual coat proteins) to the gut of the insect” (page 7, para 2, last 2 lines). The Applicnat continue to argue that “there is nothing in Fuentes that remedies the deficiencies of Bonning and Zhou noted above” (page 7, para 3, line 4-5). Applicant argues that Fuentes reports silencing of mRNA encoding the TYLCV CP via RNAi to prevent TYLCV virion production, whereas the present invention utilizes the TYLCV CP to deliver insect toxins to the gut of whiteflies (p. 7, 3rd paragraph).
The Applicant also requests to hold the double patenting rejection in abeyance until the is an indication of allowable subject matter (page 8, para 1, last 2 lines).
The Examiner disagrees. Bonning et al. describes that coat proteins of an insect vectored plant viruses can be used to deliver toxins into the hemocoel (page 11, para 0111, line 11-14). Bonning et al. also teaches that once delivered, the insect specific toxins either act at the gut epithelium or act within the hemocoel (body cavity). That structural feature is/was not restricted to JcDNV but also present in many viruses from different families including Luteovirus (Bonning et al., page 12, para 0113, last 3 lines). The only required feature needed by the (coat) protein is whether the protein can reach target destination, i.e., insect gut epithelium and/or hemocoel.
Zhou et al. describes that TYLCV CP is the only structural protein known to be involved in the virus transmission (page 1, right column, para 2). Once acquired orally by whitefly, TYLCV CP follows a sequential path of stylet-midgut-haemolymph-salivary gland, and in this journey inside whitefly, virus has to cross two major physiological barriers, namely midgut and salivary gland, to be transmitted successfully by vector (page 1, left column, para 1; Fig. 5), implying that TYLCV CP can pass across the gut epithelium and moves into the hemocoel of the insect vector (i.e., whitefly). These two teachings make it obvious to an ordinarily skilled artisan and would have motivated the artisan to express a fusion protein comprising an insect specific toxin and the TYLCV CP which were known to pass across the gut epithelium and move into the hemocoel of the target insect pest, as discussed above.
The Applicant does not provide any evidence that TYLCV CP (in absence of entire virion) would have behaved differently in terms of reaching the gut and/or hemocoel of its target insect (a whitefly). Applicant’s opinion cannot take the place of evidence (MPEP 716.01(c)(II), 2145(I)).
Regarding Applicant’s arguments in regards to Fuentes, it is noted that the reference of Fuentes et al. is cited in the rejection only for TYLCV CP sequence.
Applicant is reminded that the test for obviousness is not whether the features of a secondary reference may be bodily incorporated into the structure of the primary reference; nor is it that the claimed invention must be expressly suggested in any one or all of the references. Rather, the test is what the combined teachings of the references would have suggested to those of ordinary skill in the art. In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981).
Regarding Applicant’s request to that the double patenting rejection be held in abeyance: the double patenting rejection is maintained herewith.
Conclusion
No claim is allowed.
Communication
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAY CHATTERJEE whose telephone number is (703)756-1329. The examiner can normally be reached (Mon - Fri) 8.30 am to 5.30 pm..
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Jay Chatterjee
Patent Examiner
Art Unit 1662
/Jay Chatterjee/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
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