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1. (WO2019049913) MODIFIED CAS9 PROTEIN, AND USE THEREOF
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Pub. No.: WO/2019/049913 International Application No.: PCT/JP2018/032948
Publication Date: 14.03.2019 International Filing Date: 05.09.2018
IPC:
C12N 15/09 (2006.01) ,C12N 9/22 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9
Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14
Hydrolases (3.)
16
acting on ester bonds (3.1)
22
Ribonucleases
Applicants:
国立大学法人 東京大学 THE UNIVERSITY OF TOKYO [JP/JP]; 東京都文京区本郷七丁目3番1号 3-1, Hongo 7-chome, Bunkyo-ku, Tokyo 1138654, JP
エディジーン株式会社 EDIGENE CORPORATION [JP/JP]; 東京都中央区八丁堀2-30-16 2-30-16, Hatchobori, Chuou-ku, Tokyo 1040032, JP
Inventors:
濡木 理 NUREKI, Osamu; JP
西増 弘志 NISHIMASU, Hiroshi; JP
平野 央人 HIRANO, Hisato; JP
梶本 祥平 KAJIMOTO, Shohei; JP
山形 哲也 YAMAGATA, Tetsuya; US
シン ユアンバウ QIN, Yuanbo; US
コノリー キース エム CONNOLLY, Keith M.; US
トンプソン イーアン THOMPSON, Iain; US
Agent:
高島 一 TAKASHIMA, Hajime; JP
鎌田 光宜 KAMADA, Mitsunori; JP
Priority Data:
62/554,22705.09.2017US
62/668,96809.05.2018US
62/724,98130.08.2018US
Title (EN) MODIFIED CAS9 PROTEIN, AND USE THEREOF
(FR) PROTÉINE CAS9 MODIFIÉE ET UTILISATION DE CELLE-CI
(JA) 改変されたCas9タンパク質及びその用途
Abstract:
(EN) Provided is a mutant SaCas9 protein having the following amino acid sequence mutations in SEQ. ID NO: 2: at position 782, glutamic acid to lysine (E782K); at position 800, leucine to arginine (L800R); at position 968, asparagine to arginine (N968R); at position 985, asparagine to alanine (N985A); at position 991, arginine to alanine (R991A); at position 1021, alanine to serine (A1021S); at position 927, threonine to lysine (T927K); at position 929, lysine to asparagine (K929N); at position 1017, isoleucine to phenylalanine (I1017F). Said mutant SaCas9 protein maintains the ability to bind with guide RNA while target sequence restrictions are relaxed, and is useful as a tool for gene editing.
(FR) L'invention concerne une protéine SaCas9 mutante ayant les mutations de séquence d'acides aminés suivantes dans la SEQ. ID NO : 2 : en position 782, acide glutamique en lysine (E782K) ; en position 800, leucine en arginine (L800R) ; en position 968, asparagine en arginine (N968R) ; en position 985, asparagine en alanine (N985A) ; en position 991, arginine en alanine (R991A) ; en position 1021, alanine en sérine (A1021S) ; en position 927, thréonine en lysine (T927K) ; en position 929, lysine en asparagine (K929N) ; en position 1017, isoleucine en phénylalanine (I1017F). Ladite protéine SaCas9 mutante maintient la capacité de se lier à l'ARN guide tandis que les restrictions de séquence cible sont relâchées, et est utile en tant qu'outil pour l'édition génique.
(JA) 配列番号2において、782位のグルタミン酸がリジンに(E782K)、800位のロイシンがアルギニンに(L800R)、968位のアスパラギンがアルギニンに(N968R)、985位のアスパラギンがアラニンに(N985A)、991位のアルギニンがアラニンに(R991A)、1021位のアラニンがセリンに(A1021S)、927位のスレオニンがリジンに(T927K)、929位のリジンがアスパラギンに(K929N)、1017位のイソロイシンがフェニルアラニンに(I1017F)変異したアミノ酸配列を含むタンパク質等の変異型SaCas9タンパク質は、ガイドRNAとの結合能を維持しつつ標的配列の制限が緩和されており、遺伝子編集におけるツールとして有用である。
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Japanese (JA)
Filing Language: Japanese (JA)