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 .
Election/Restrictions
Applicant’s election of Group I in the reply filed on 10/08/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
The Applicant also elects the following species without traverse:
SEQ ID NO: 25, in claim 2
SEQ ID NO: 4, in claim 3
MCP, in claim 5
SEQ ID NO: 13, in claim 7
SEQ ID NO: 13, in claim 8
APOBEC1 deaminase, in claim 14
SEQ ID NO: 29, in claim 16
However, SEQ ID NO: 26 is rejoined.
The election is made FINAL.
Claim Status
Claims 1-36 are pending.
Claims 28-36 are withdrawn from examination as being part of nonelected invention.
Claims 1-27 are being examined.
Claim Objections
Claims 2 and 12 are objected to because of the following informalities:
Claim 2 recites “Ca9 nickase” in line 1. It appears that the Applicant implies “Cas9 nickase”
Claim 12, recites, “the second scan” in line 2. It appears that the Applicant implies “scRNA”.
Appropriate correction is required.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 2, 14-15, 19 and 21-23 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claims 2, 19 and 21-22; the phrase "for example" renders the claim indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Regarding claims 14-15, the claims recite, “… functional variant(s)…” of cytosine deaminases comprising APOBEC1 deaminase, activation-induced cytidine deaminase (AID), APOBEC3G, CDA1, human APOBEC3A deaminase. The Applicant does not define the term. It is not clear what the Applicant imply by “functional variants”. Do such “variants” include any deaminase including adenine deaminase found in any organism including bacteria and fungus, for example? As recited in the claims, the term is also not limited to any percentage of homology/identity to any specific SEQ ID NO. Therefore, the metes and bounds of the recited expression of “functional variant(s)” is not clear in the claims.
Regarding claims 23 the phrase "preferably" renders the claim indefinite because use of a narrow range that falls within a broader range in the same claim may render the claim indefinite when the boundaries of the claim are not discernible. See MPEP § 2173.05(c).
Claim Rejections - 35 USC § 112(a)
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 7-9 and 12-15 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 claims contain 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.
The Applicant describes, “studies have demonstrated that scRNA formed by adding two MS2 hairpins to the 3′ end of (e)sgRNA can efficiently mediate CRISPRa in human cells, wherein MS2 is a commonly used RNA aptamer. Therefore, the scRNA vector pOsU3-esgRNA-2×MS2 driven by the OsU3 promoter was first constructed” (page 19, line 25-28). Such “scaffold system” of gene editing using scRNA is known in the art (Shakirova et al., Cell Reprogramming With CRISPR/Cas9 Based Transcriptional Regulation Systems, 2020, Front. Bioeng. Biotechnol., 8:882; page 4, left column, para 6, line 1-8). The sequences of 2xMS2 aptamer and the (MS2) aptamer-specific binding protein MCP are standard and long well known in the art ((Zalatan et al. (US 2017/0233762 A1) describes 2xMS2 aptamer sequence (SEQ ID NO: 9) and MCP protein sequence (SEQ ID NO: 2)).
Regarding claims 7-9, following the description of the instant invention and fusing the 2xMS aptamer sequence (small letters below) to the 3’end of the gRNA (SEQ ID NO: 4) (capital letters below) would result in the following sequence: GTTTAAGAGCTATGCTGGAAACAGC ATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTGTTTTTTATGTCT gggagcacatgaggatcacccatgtgccacgagcgacatgaggatcacccatg tcgctcgtgttccc. Sequence alignment of the fused sequence with instant SEQ ID NO: 13 shows an extra sequence, as highlighted in grey, between the gRNA (SEQ ID NO: 4) and the 2xMS2 sequence, as shown below.
Title: US-17-909-309A-13
Perfect score: 173
Sequence: 1 gtttaagagctatgctggaa..........tttttttgttttttatgtct 173
Searched: 1 seqs, 172 residues
Database : NASEQ2_10222025_121824.seq:*
RESULT 1
NASEQ2_10222025_121824
Query Match 71.7%; Score 124; DB 1; Length 172; Best Local Similarity 89.0%;
Matches 153; Conservative 0; Mismatches 0; Indels 19; Gaps 1;
Qy 1 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC 60
Qy 61 TTGAAAAAGTGGCACCGAGTCGGTGC-------------------GGGAGCACATGAGGA 101
|||||||||||||||||||||||||| |||||||||||||||
Db 61 TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTGTTTTTTATGTCTGGGAGCACATGAGGA 120
Qy 102 TCACCCATGTGCCACGAGCGACATGAGGATCACCCATGTCGCTCGTGTTCCC 153
||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TCACCCATGTGCCACGAGCGACATGAGGATCACCCATGTCGCTCGTGTTCCC 172
The Applicant does not describe if the grey highlighted sequence in critical in the context of the invention. The Applicant also does not describe where the grey highlight sequence came from and why it is used while performing the function of editing the plant genome. A skilled artisan would not be able to design a suitable scRNA to edit a target genome as provided in the instant description.
Regarding claims 12-13, sequence alignment between instant SEQ ID NO: 22 (as recited in claim 12) and the (second) gRNA comprising the sequence ID NO: 3 (as recited in claim 3) fused with the 2xMS2 sequence (as described above) is shown below.
CLUSTAL 2.1 multiple sequence alignment
SEQ22 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC
2xMS2+gRNA3 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC
************************************************************
SEQ22 TTGAAAAAGTGGCACCGAGTCGGTGCG---------------------------------
2xMS2+gRNA3 TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTGTTTTTTATGTCTGTTTTAGAGCTAGA
**************************
SEQ22 -GGAGCGCCCTGAAGAAGGGCG---CCTGCTGCGGCCCTGAAGAAGGGCCGCAGCAGTTC
2xMS2+gRNA3 AATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGT
. ***.. *.**.:*.*** ** : *..* .**.*. ***. *.. :
SEQ22 CCTTTTTTTGTTTTTTATGTCT
2xMS2+gRNA3 GCTTTTTTTGTTTTTTATGTCT
*********************
In this case, the grey highlighted (TTTTTTGTTTTTTATGTCT) sequence (which does not arise from either the 2xMS2 sequence or the gRNA sequence) is also present. Moreover, in this case, the (second) gRNA sequence comprising instant SEQ ID NO: 3 is not aligning with the second scRNA comprising SEQ ID NO: 22. Further, the second RNA aptamer specific binding protein comprising SEQ ID NO: 36 (as recited in claim 13) is not MCP and would not bind to the 2xMS2 sequence present in the scRNA comprising the sequence of SEQ ID NO: 22.
Regarding claims 14-15, the instant description teaches several examples of specific cytosine deaminases comprising APOBEC1 deaminase, activation-induced cytidine deaminase (AID), APOBEC3G, CDA1, and human APOBEC3A deaminase. However, the Applicant does not describe or define the term “functional variant(s)” of cytosine deaminase.
An invention described solely in terms of a method of making and/or its function would lack written descriptive support where there is no described (in the specification) or art-recognized correlation between the disclosed function and the structure(s) responsible for the function. See MPEP § 2163.
Considering the breadth of the claims and lack of structure function relationship based on the instant description, the Applicant does not appear to have been in possession of the claimed genus at the time this application was filed.
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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 4-6, 14-15 and 20 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Brezgin et al. (Dead Cas Systems: Types, Principles, and Applications, 2019, Int. J. Mol. Sci., 20, 6041).
Claim 1 is drawn to a multiplex genome editing system in a plant comprising a CRISPR nickase along with one or more scRNAs wherein each scRNA contains an aptamer, a fusion protein that comprises an aptamer-specific binding protein, and a nucleotide base (adenine or cytosine) deamination domain.
Brezgin et al. describes a multiplex genome editing system in various organisms including plants (page 1, para 1, line 3) using nickase nCas9 (nCas9) which increases efficiency of base editing (page 12, para 6). Brezgin et al. also describes the “scaffold technique” (page 7, Fig. 2) where modified gRNAs comprising scaffold RNA (scRNA) containing two MS2 aptamers (as recited in claims 4 and 6) (page 3, para 9, line 5-6) and an aptamer-specific binding protein (MCP) (as recited in claim 5) fused to effector molecule(s) resulting in fusion proteins (of aptamer-specific binding proteins and effector proteins). Effectors proteins can be a deaminase, either for adenine or cytosine.
It is well known in the art that aptamers are part of scRNAs (i.e., the scaffold sequence in a gRNA). Brezgin et al. describes simultaneously editing or modulating many genes to control complex biological processes with unprecedented accuracy. This multiplexing is done by a Cas endonuclease that cuts a single RNA transcript into many gRNAs (i.e., paired gRNAs) which would read on to targeting individual targets (page 15, para 4), which can be more than three, and would read on to “first”, “second”, and “third” scRNA and corresponding “first”, “second”, and “third” target regions, as recited in claim 1. Brezgin et al. describes various effector proteins comprising cytidine/cytosine deaminases like APOBEC1 (page 13, para 2, line 1) (as recited in claims 14-15); and adenine deaminases including TadA adenosine deaminase (page 12, para 5, last 2 lines) and ADAR2 adenosine deaminase (page 13, para 4), which are known to comprise adenine deamination domain. TadA is a DNA dependent adenine deaminase (page 12, para 5, line 9-10), as recited in claim 20, that specifically targets DNA (page 15, table 4).
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 2-3 and 21-26 are rejected under 35 U.S.C. 103 as being unpatentable over Brezgin et al. as applied to claims 1, 4-6, 14-15 and 20 above (see rejections under 35 USC §102), and further in view of Gao et al. (WO2019120283A1).
Claim 2 is drawn to a Cas9 nickase comprising the amino acid sequence of SEQ ID NO: 25, while claim 3 is drawn to a gRNA sequence having 100% sequence identity to instant SEQ ID NO: 3.
Brezgin et al. describes a multiplex genome editing system comprising a Cas9 nickase (nCas9) along with one or more scRNAs wherein each scRNA contains an aptamer, a fusion protein that comprises an aptamer-specific binding protein, and a nucleotide base (adenine or cytosine) deamination domain, as described above. Brezgin et al. also describes use of peptide linkers (as recited in claims 23 and 25) to further enhance efficacy of the fusion proteins (page 3, para 6, line 3-5; page 4, para 4, line 3-5)
However, Brezgin et al. does not describe a Cas9 nickase comprising the amino acid sequence of SEQ ID NO: 25 or a polynucleotide sequence set forth in SEQ ID NO: 4.
Gao et al. describes a method for performing efficient base editing to a target sequence in a plant genome by a Cas9-cytidine deaminase fusion protein. It describes simultaneous editing three homoalleles in hexaploid bread wheat conferring heritable resistance to powdery mildew (page 23, para 2, line 2-4). All the proteins including the specific fusion proteins needed for base editing especially for editing genomic DNA need to be present in the nucleus in the cell. Gao et al. describes Nuclear Localization Sequence (NLS) (page 15, line 1-3), as recited in claim 26. It teaches one or more NLSs in the base-editing fusion proteins that are sufficient to drive the base-editing fusion protein into the nucleus of a plant cell to achieve base editing (page 15, line 3-6).
Gao et al. teaches a nuclease-inactivated Cas9 nickase (SEQ ID NO: 3) (page 5, para 0060) comprising 100% identity to instant SEQ ID NO: 25, as shown below.
RESULT 1
BGM50953
ID BGM50953 standard; protein; 1367 AA.
AC BGM50953;
DT 22-AUG-2019 (first entry)
DE Streptococcus pyogenes Cas9 protein, SEQ ID 3.
KW CRISPR associated 9; CRISPR-Cas9 system; Cas9 protein; crop improvement;
KW genome editing; herbicide resistance; plant breeding; transgenic plant.
OS Streptococcus pyogenes.
CC PN WO2019120283-A1.
CC PD 27-JUN-2019.
CC PF 21-DEC-2018; 2018WO-CN122640.
PR 21-DEC-2017; 2017CN-11393160.
PR 28-APR-2018; 2018CN-10402244.
CC PA (CAGD ) INST GENETICS & DEV BIOL CAS.
CC PI Gao C, Li C, Zong Y, Wang Y;
DR WPI; 2019-53605Q/53.
CC PT System useful for base editing target sequence in plant genome, comprises base-editing fusion protein, and guide RNA, and expression construct comprising nucleotide sequence encoding base-editing fusion protein, and guide RNA.
CC PS Claim 4; SEQ ID NO 3; 60pp; English.
CC PT System useful for base editing target sequence in plant genome, comprises
base-editing fusion protein, and guide RNA, and expression construct comprising nucleotide sequence encoding base-editing fusion protein, and guide RNA.
CC PS Example 5; Fig 7; 60pp; English.
CC The present invention relates to a novel system useful for base editing of a target sequence in a plant genome. The system comprises: base-editing fusion protein, and a guide RNA; an expression construct comprising a nucleotide sequence encoding a base-editing fusion protein, and a guide RNA; a base-editing fusion protein corresponds to SEQ ID NO: 4, 5 or 18-25 or 33-39 (see BGM50954-BGM50955 or BGM50968-BGM50975 or BGM50983-BGM50989), and an expression construct comprising a nucleotide sequence encoding the guide RNA; an expression construct comprising a nucleotide sequence corresponds to SEQ ID NO: 6, 7 or 10-17 or 26-32 (see BGM50956-BGM50957 or BGM50960-BGM50967 or BGM50976-BGM50982) encoding the base-editing fusion protein, and an expression construct comprising a nucleotide sequence encoding the guide RNA; and an expression construct comprising a nucleotide sequence encoding the base-editing fusion protein and a nucleotide sequence encoding the guide RNA, where the base-editing fusion protein comprises a nuclease-inactivated clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated 9 (Cas9) effector protein corresponds to SEQ ID NO: 3 (see BGM50953) and a DNA-dependent adenine deaminase (tRNA adenine deaminase or TadA) corresponds to SEQ ID NO: 1 (see BGM50951). The invention further relates to: (1) a method for producing a genetically modified plant; (2) a genetically modified plant or its progeny, or a part, where the plant; and (3) a method for plant breeding. The system of the invention is also used for producing a plant having herbicide resistance.
Query Match 100.0%; Score 6998; Length 1384; Best Local Similarity 100.0%;
Matches 1367; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2 DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA 61
Qy 61 TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 62 TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN 121
Qy 121 IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 122 IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV 181
Qy 181 DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 182 DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL 241
Qy 241 IALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 242 IALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 301
Qy 301 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 302 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG 361
Qy 361 YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 362 YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA 421
Qy 421 ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 422 ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV 481
Qy 481 VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 482 VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 541
Qy 541 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 542 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII 601
Qy 601 KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGR 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 602 KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGR 661
Qy 661 LSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLH 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 662 LSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLH 721
Qy 721 EHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 722 EHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM 781
Qy 781 KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHI 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 782 KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHI 841
Qy 841 VPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 842 VPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLT 901
Qy 901 KAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSK 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 902 KAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSK 961
Qy 961 LVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKM 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 962 LVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKM 1021
Qy 1021 IAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1022 IAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFA 1081
Qy 1081 TVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAY 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1082 TVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAY 1141
Qy 1141 SVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKY 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1142 SVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKY 1201
Qy 1201 SLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQ 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1202 SLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQ 1261
Qy 1261 HKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAP 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1262 HKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAP 1321
Qy 1321 AAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 1367
|||||||||||||||||||||||||||||||||||||||||||||||
Db 1322 AAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 1368
Regarding claim 3, Gao et al. describes an gRNA having 100% sequence identity to instant SEQ ID NO: 4, as shown below.
RESULT 3
BGM51116
ID BGM51116 standard; DNA; 487 BP.
AC BGM51116;
DT 22-AUG-2019 (first entry)
DE Wheat U6 promoter target sequence (pTaU6-esgRNA).
KW CRISPR-Cas9 system; U6 gene; crop improvement; ds; genome editing; herbicide resistance; plant; plant breeding; transgenic plant.
OS Triticum aestivum.
OS Synthetic.
CC PN WO2019120283-A1.
CC PD 27-JUN-2019.
CC PF 21-DEC-2018; 2018WO-CN122640.
PR 21-DEC-2017; 2017CN-11393160.
PR 28-APR-2018; 2018CN-10402244.
CC PA (CAGD ) INST GENETICS & DEV BIOL CAS.
CC PI Gao C, Li C, Zong Y, Wang Y;
DR WPI; 2019-53605Q/53.
CC PT System useful for base editing target sequence in plant genome, comprises
base-editing fusion protein, and guide RNA, and expression construct comprising nucleotide sequence encoding base-editing fusion protein, and guide RNA.
CC PS Example 5; Fig 7; 60pp; English.
The present invention relates to a novel system useful for base editing of a target sequence in a plant genome. The system comprises: base-editing fusion protein, and a guide RNA; an expression construct comprising a nucleotide sequence encoding a base-editing fusion protein, and a guide RNA; a base-editing fusion protein corresponds to SEQ ID NO: 4, 5 or 18-25 or 33-39 (see BGM50954-BGM50955 or BGM50968-BGM50975 or BGM50983-BGM50989), and an expression construct comprising a nucleotide sequence encoding the guide RNA; an expression construct comprising a nucleotide sequence corresponds to SEQ ID NO: 6, 7 or 10-17 or 26-32 (see BGM50956-BGM50957 or BGM50960-BGM50967 or BGM50976-BGM50982) encoding the base-editing fusion protein, and an expression construct comprising a nucleotide sequence encoding the guide RNA; and an expression construct comprising a nucleotide sequence encoding the base-editing fusion protein and a nucleotide sequence encoding the guide RNA, where the base-editing fusion protein comprises a nuclease-inactivated clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR associated 9 (Cas9) effector protein corresponds to SEQ ID NO: 3 (see BGM50953) and a DNA-dependent adenine deaminase (tRNA adenine deaminase or TadA) corresponds to SEQ ID NO: 1 (see BGM50951). The invention further relates to: (1) a method for producing a genetically modified plant; (2) a genetically modified plant or its progeny, or a part, where the plant; and (3) a method for plant breeding. The system of the invention is also used for producing a plant having herbicide resistance.
SQ Sequence 487 BP; 116 A; 110 C; 125 G; 136 T; 0 U; 0 Other;
Query Match 100.0%; Score 106; Length 487; Best Local Similarity 100.0%;
Matches 106; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 382 GTTTAAGAGCTATGCTGGAAACAGCATAGCAAGTTTAAATAAGGCTAGTCCGTTATCAAC 441
Qy 61 TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTGTTTTTTATGTCT 106
||||||||||||||||||||||||||||||||||||||||||||||
Db 442 TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTGTTTTTTATGTCT 487
Regarding claims 21-23, Gao et al. describes a TadA DNA-dependent adenine deaminase (SEQ ID NO: 2) (page 13, line 14-16) comprising 100% sequence identity to instant SEQ ID NO: 33, as recited in claim 21, as shown below.
RESULT 1
BGM50952
ID BGM50952 standard; protein; 166 AA.
AC BGM50952;
DT 22-AUG-2019 (first entry)
DE Escherichia coli tRNA adenine deaminase (TadA) variant, SEQ ID 2.
KW CRISPR-Cas9 system; DNA-dependent adenine deaminase; TadA protein;
KW crop improvement; genome editing; herbicide resistance; mutein;
KW plant breeding; tRNA adenine deaminase; transgenic plant.
OS Escherichia coli.
FT /note= "Wild type Trp is substituted by Arg"
FT Misc-difference 35
FT /note= "Wild type His is substituted by Leu"
FT Misc-difference 47
FT /note= "Wild type Pro is substituted by Ala"
FT Misc-difference 50
FT /note= "Wild type Arg is substituted by Leu"
FT Misc-difference 83
FT /note= "Wild type Leu is substituted by Phe"
FT Misc-difference 105
FT /note= "Wild type Arg is substituted by Val"
FT Misc-difference 107
FT /note= "Wild type Asp is substituted by Asn"
FT Misc-difference 122
FT /note= "Wild type His is substituted by Tyr"
FT Misc-difference 145
FT /note= "Wild type Ser is substituted by Cys"
FT Misc-difference 146
FT /note= "Wild type Asp is substituted by Tyr"
FT Misc-difference 151
FT /note= "Wild type Arg is substituted by Pro"
FT Misc-difference 154
FT /note= "Wild type Glu is substituted by Val"
FT Misc-difference 155
FT /note= "Wild type Ile is substituted by Phe"
FT Misc-difference 156
FT /note= "Wild type Lys is substituted by Asn"
CC PN WO2019120283-A1.
CC PD 27-JUN-2019.
CC PF 21-DEC-2018; 2018WO-CN122640.
PR 21-DEC-2017; 2017CN-11393160.
PR 28-APR-2018; 2018CN-10402244.
CC PA (CAGD ) INST GENETICS & DEV BIOL CAS.
CC PI Gao C, Li C, Zong Y, Wang Y;
DR WPI; 2019-53605Q/53.
CC PT System useful for base editing target sequence in plant genome, comprises
CC PT base-editing fusion protein, and guide RNA, and expression construct
CC PT comprising nucleotide sequence encoding base-editing fusion protein, and
CC PT guide RNA.
CC PS Claim 3; SEQ ID NO 2; 60pp; English.
CC The present invention relates to a novel system useful for base editing
CC of a target sequence in a plant genome. The system comprises: base-
CC editing fusion protein, and a guide RNA; an expression construct
CC comprising a nucleotide sequence encoding a base-editing fusion protein,
CC and a guide RNA; a base-editing fusion protein corresponds to SEQ ID NO:
CC 4, 5 or 18-25 or 33-39 (see BGM50954-BGM50955 or BGM50968-BGM50975 or
CC BGM50983-BGM50989), and an expression construct comprising a nucleotide
CC sequence encoding the guide RNA; an expression construct comprising a
CC nucleotide sequence corresponds to SEQ ID NO: 6, 7 or 10-17 or 26-32 (see
CC BGM50956-BGM50957 or BGM50960-BGM50967 or BGM50976-BGM50982) encoding the
CC base-editing fusion protein, and an expression construct comprising a
CC nucleotide sequence encoding the guide RNA; and an expression construct
CC comprising a nucleotide sequence encoding the base-editing fusion protein
CC and a nucleotide sequence encoding the guide RNA, where the base-editing
CC fusion protein comprises a nuclease-inactivated clustered regularly
CC interspaced short palindromic repeat (CRISPR)-CRISPR associated 9 (Cas9)
CC effector protein corresponds to SEQ ID NO: 3 (see BGM50953) and a DNA-
CC dependent adenine deaminase (tRNA adenine deaminase or TadA) corresponds
CC to SEQ ID NO: 1 (see BGM50951). The invention further relates to: (1) a
CC method for producing a genetically modified plant; (2) a genetically
CC modified plant or its progeny, or a part, where the plant; and (3) a
CC method for plant breeding. The system of the invention is also used for
CC producing a plant having herbicide resistance. Note: The mutation
CC positions for this sequence as given in the claim 2 of the specification
CC do not correctly match. Therefore, the mutation position given in the
CC feature table have been deduced by comparing this sequence with the
CC parent sequence (see BGM50951).
SQ Sequence 166 AA;
Query Match 100.0%; Score 869; Length 166; Best Local Similarity 100.0%;
Matches 166; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIM 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIM 60
Qy 61 ALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVL 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVL 120
Qy 121 HYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTD 166
||||||||||||||||||||||||||||||||||||||||||||||
Db 121 HYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTD 166
Gao et al. describes an adenine base editing system comprising SEQ ID NO: 24, which is suitable for plant cell editing (page 27, line 10-18), besides a DNA dependent adenine deaminase from E. coli (page 12, line 11-14), as recited in claim 23. SEQ ID NO: 24 of Gao et al. is codon optimized for plant expression and having 100% sequence identity to instant SEQ ID NO: 32 (as recited in claim 22), as shown below.
RESULT 31
BGM50974
ID BGM50974 standard; protein; 809 AA.
AC BGM50974;
DT 22-AUG-2019 (first entry)
DE E. coli TadA-Cas9 fusion protein (SanCas9-ABE-1), SEQ ID 24.
KW CRISPR associated 9; CRISPR-Cas9 system; Cas9 protein;
KW DNA-dependent adenine deaminase; TadA protein; chimeric protein;
KW crop improvement; fusion protein; genome editing; herbicide resistance;
KW plant breeding; tRNA adenine deaminase; transgenic plant.
OS Escherichia coli.
CC PN WO2019120283-A1.
CC PD 27-JUN-2019.
CC PF 21-DEC-2018; 2018WO-CN122640.
PR 21-DEC-2017; 2017CN-11393160.
PR 28-APR-2018; 2018CN-10402244.
CC PA (CAGD ) INST GENETICS & DEV BIOL CAS.
CC PI Gao C, Li C, Zong Y, Wang Y;
DR WPI; 2019-53605Q/53.
DR N-PSDB; BGM50966.
CC PT System useful for base editing target sequence in plant genome, comprises base-editing fusion protein, and guide RNA, and expression construct comprising nucleotide sequence encoding base-editing fusion protein, and guide RNA.
CC PS Claim 10; SEQ ID NO 24; 60pp; English.
CC The present invention relates to a novel system useful for base editing
CC of a target sequence in a plant genome. The system comprises: base-
CC editing fusion protein, and a guide RNA; an expression construct
CC comprising a nucleotide sequence encoding a base-editing fusion protein,
CC and a guide RNA; a base-editing fusion protein corresponds to SEQ ID NO:
CC 4, 5 or 18-25 or 33-39 (see BGM50954-BGM50955 or BGM50968-BGM50975 or
CC BGM50983-BGM50989), and an expression construct comprising a nucleotide
CC sequence encoding the guide RNA; an expression construct comprising a
CC nucleotide sequence corresponds to SEQ ID NO: 6, 7 or 10-17 or 26-32 (see
CC BGM50956-BGM50957 or BGM50960-BGM50967 or BGM50976-BGM50982) encoding the
CC base-editing fusion protein, and an expression construct comprising a
CC nucleotide sequence encoding the guide RNA; and an expression construct
CC comprising a nucleotide sequence encoding the base-editing fusion protein
CC and a nucleotide sequence encoding the guide RNA, where the base-editing
CC fusion protein comprises a nuclease-inactivated clustered regularly
CC interspaced short palindromic repeat (CRISPR)-CRISPR associated 9 (Cas9)
CC effector protein corresponds to SEQ ID NO: 3 (see BGM50953) and a DNA-
CC dependent adenine deaminase (tRNA adenine deaminase or TadA) corresponds
CC to SEQ ID NO: 1 (see BGM50951). The invention further relates to: (1) a
CC method for producing a genetically modified plant; (2) a genetically
CC modified plant or its progeny, or a part, where the plant; and (3) a
CC method for plant breeding. The system of the invention is also used for
CC producing a plant having herbicide resistance.
SQ Sequence 809 AA;
Query Match 100.0%; Score 872; Length 809; Best Local Similarity 100.0%;
Matches 166; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 SEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEIM 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2 SEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEIM 61
Qy 61 ALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 62 ALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL 121
Qy 121 HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTD 166
||||||||||||||||||||||||||||||||||||||||||||||
Db 122 HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTD 167
Before the effective filing date, it would have been obvious to an ordinarily skilled artisan to use Cas9 nickase or a functional equivalent of a nuclease inactivated Cas9 nickase, and a gRNA to edit target gene(s)/allele(s) in a plant, as described by Gao et al. Designing gRNA(s) for specific purpose is a well-known standard process that depends on specific target sequence. Using a functional equivalent of Cas9 nickase, a gRNA, and a DNA-dependent adenine deaminase, as described by Gao et al., having specific structures/sequences is an experimental design choice of an ordinarily skilled artisan without having any realistic possibility of changing the outcome.
Before the effective filing date of the invention, one ordinarily skilled in the art would have been motivated to use specific Cas9 nickase, gRNA, and a DNA-dependent adenine deaminase (as described by Gao et al.) to edit target gene(s)/allele(s) in a plant.
Regarding claims 23-25, All the fused proteins (DNA or RNA dependent aptamer specific binding proteins fused with deaminase protein/domain) acts in trans in a cell. It is an experimental design choice of an ordinarily skilled artisan to make fusion proteins by fusing the aptamer-specific binding proteins (e.g. MCP) with either N-terminal or the C-terminal (as recited in claims 23-24) of an effector protein like adenine deaminase (TadA) or the corresponding adenine deamination domain (as recited in claim 24) using a linker protein. C-terminal deamination domain of TadA proteins including in E coli (as recited in claim 23) and in plants are known in the art since long1. Using a linker (as recited in claims 23 and 25) while making fusion protein(s) by fusing two or more proteins and/or domains is a standard practice in the art, because it offers several advantages such as improving biological activity, increasing expression yield, and achieving desirable pharmacokinetic profiles2 and also as described by Brezgin et al.
It is "the normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine” the exact experimental design that works more efficiently in a specific experiment, unless there is evidence indicating a specific experimental design used by the Applicant is critical. Such “critical” feature(s) need(s) to be described by the instant description. See MPEP § 2144.05.
Claims 2 and 21-26 are rejected under 35 U.S.C. 103 as being unpatentable over Brezgin et al. as applied to claims 1, 4-6, 14-15 and 20 above, and further in view of Liu et al. (WO2018027078 A1).
Brezgin et al. describes a multiplex genome editing system comprising a Cas9 nickase (nCas9) along with one or more scRNAs wherein each scRNA contains an aptamer, a fusion protein that comprises an aptamer-specific binding protein, and a nucleotide base (adenine or cytosine) deamination domain, as described above.
However, Brezgin et al. does not describe a Cas9 nickase comprising the amino acid sequence of SEQ ID NO: 25, an Escherichia coli tRNA adenine deaminase TadA (ecTadA) comprising the amino acid sequence shown in SEQ ID NO: 32 or SEQ ID NO: 33.
Liu et al. describes a method of targeted gene editing by using fusion proteins using Cas9 nickase and different adenosine deaminases that are capable of deaminating adenosine in DNA. The fusion proteins comprises one or more linkers (as recited in claim 23) (page 4, para 0009, line 6-7) and one or more nuclear localization sequences (NLS) (as recited in claim 26) (page 4, para 0009, last 2 lines).It describes a Cas9 comprising the amino acid sequence (SEQ ID NO: 35) (page , para 202; para 280) having 100% sequence identity to instant SEQ ID NO: 25, as shown below.
BLM40833
ID BLM40833 standard; protein; 167 AA.
AC BLM40833;
DT 08-SEP-2022 (first entry)
DE E. coli TadA mutant protein (pNMG-619/620/624).
KW CRISPR-Cas9 system; TadA; cytostatic; genetic disorder;
KW genetic-disease-gen.; genome editing; lysosome storage disease;
KW metabolic disorder; metabolic-gen.; neoplasm; neuroprotective;
KW recombinant protein; tRNA adenosine deaminase A; therapeutic; mutein.
OS Escherichia coli.
OS Synthetic.
CC PN WO2018027078-A1.
CC PD 08-FEB-2018.
CC PF 03-AUG-2017; 2017WO-US045381.
PR 03-AUG-2016; 2016US-0370684P.
PR 02-FEB-2017; 2017US-0454035P.
PR 20-MAR-2017; 2017US-0473714P.
CC PA (HARD ) HARVARD COLLEGE.
CC PI Gaudelli N, Liu DR;
DR WPI; 2018-11292C/13.
CC PT New adenosine deaminase capable of deaminating adenine of deoxyadenosine
CC PT in DNA useful for editing nucleobase pair of double-stranded DNA
CC PT sequence.
CC PS Claim 113; Page; 1450pp; English.
CC The invention relates to a novel adenosine deaminase capable of
CC deaminating adenine of deoxyadenosine in DNA useful for editing
CC nucleobase pair of double-stranded DNA sequence. The invention claims: 1)
CC a fusion protein; 2) a complex comprising the fusion protein and a guide
CC RNA (gRNA) bound to the napDNAbp of the fusion protein, where the
CC napDNAbp is a clustered regularly interspaced short palindromic repeat
CC (CRISPR) associated protein 9 (Cas9) domain, a Cpfl, a CasX, a CasY, a
CC C2c1, a C2c2, or a C2c3.; 3) a method involving (a) contacting a nucleic
CC acid molecule with the fusion protein and a gRNA, where the gRNA is 15-
CC 100 nucleotides long and comprises a sequence of at least 10 contiguous
CC nucleotides that is complementary to a target sequence, or (b) contacting
CC a nucleic acid molecule with the complex; 4) a method for editing a
CC nucleobase pair of a double-stranded DNA (dsDNA) sequence; 5) a nucleic
CC acid-guided adenosine deaminase coupled to an inhibitor of base excision
CC repair; 6) a kit comprising a nucleic acid construct; 7) a polynucleotide
CC encoding the adenosine deaminase, or the fusion protein; 8) a vector
CC comprising the polynucleotide; and 9) a cell comprising (a) the adenosine
CC deaminase, or the fusion protein, (b) the complex, or (c) the nucleic
CC acid molecule encoding the adenosine deaminase, or the fusion protein.
CC The adenosine deaminase is used in pharmaceutical composition for editing
CC a nucleobase pair of a dsDNA sequence; for modifying a polynucleotide,
CC and treating a proliferative disease, genetic disease, neoplastic
CC disease, metabolic disease, and lysosomal storage disease. The adenosine
CC deaminase has excellent stability. The present sequence represents an
CC Escherichia coli tRNA adenosine deaminase A (TadA) W23R/H36L/P48A/R51L/L8
CC 4F/A106V/D108N/H123Y/S146C/D147Y/R152P/E155V/I156F/K157N mutant protein
CC (pNMG-619 / pNMG-620 / pNMG-624) used for editing nucleobase pair of
CC double-stranded DNA sequence. Note: The present sequence is not shown in
CC the specification but is created based on the information given in table
CC 4 of the specification from the E. coli TadA protein shown as SEQ ID NO:
CC 1 (see BFB10528).
SQ Sequence 167 AA;
Query Match 100.0%; Score 6998; Length 1367;
Best Local Similarity 100.0%;
Matches 1367; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA 60
Qy 61 TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN 120
Qy 121 IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV 180
Qy 181 DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNL 240
Qy 241 IALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 IALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 300
Qy 301 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG 360
Qy 361 YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA 420
Qy 421 ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV 480
Qy 481 VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 540
Qy 541 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII 600
Qy 601 KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGR 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGR 660
Qy 661 LSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLH 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 LSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLH 720
Qy 721 EHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 EHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM 780
Qy 781 KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHI 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHI 840
Qy 841 VPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 VPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLT 900
Qy 901 KAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSK 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 KAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSK 960
Qy 961 LVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKM 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 LVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKM 1020
Qy 1021 IAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFA 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 IAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFA 1080
Qy 1081 TVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAY 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 TVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAY 1140
Qy 1141 SVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYK