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1. (WO2019046636) DOUBLE SELECTION HDR CRISPR-BASED EDITING
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Pub. No.: WO/2019/046636 International Application No.: PCT/US2018/048943
Publication Date: 07.03.2019 International Filing Date: 30.08.2018
Chapter 2 Demand Filed: 21.12.2018
IPC:
A61K 35/34 (2015.01) ,C12N 5/074 (2010.01) ,C12N 9/12 (2006.01) ,C12N 15/90 (2006.01)
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
K
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
35
Medicinal preparations containing material or reaction products thereof with undetermined constitution
12
Materials from mammals or birds
34
Muscles; Heart
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
5
Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
07
Animal cells or tissues
071
Vertebrate cells or tissues, e.g. human cells or tissues
074
Adult stem cells
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
10
Transferases (2.)
12
transferring phosphorus containing groups, e.g. kinases (2.7)
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
87
Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
90
Stable introduction of foreign DNA into chromosome
Applicants:
CLAUSSNITZER, Melina [US/US]; US
GOGGIN, Sarah; US
SAADAT, Alham; US
THE BROAD INSTITUTE, INC. [US/US]; 415 Main Street Cambridge, Massachusetts 02142, US
Inventors:
CLAUSSNITZER, Melina; US
GOGGIN, Sarah; US
SAADAT, Alham; US
Agent:
REY, Gertrud U.; US
NIX, F. Brent; US
Priority Data:
62/552,33330.08.2017US
Title (EN) DOUBLE SELECTION HDR CRISPR-BASED EDITING
(FR) ÉDITION BASÉE SUR CRISPR DE TYPE HDR À DOUBLE SÉLECTION
Abstract:
(EN) The invention provides homology directed repair (HDR) constructs for variant screening in cells comprising: a left and right homology arm, with either the left or right homology arm encoding a genomic edit to be incorporated at a target locus; and an excisable double selection cassette located within the left and right homology arms, the excisable double selection cassette comprising; a first selection marker; and a second selection marker; and wherein the first selection marker and the second selection marker are located between a first and second excision site. Also provided are homology directed repair (HDR) vectors comprising a construct as described herein, and methods for using such vectors.
(FR) L'invention concerne des constructions de réparation dirigée par homologie (HDR) pour identifier par criblage des variants dans des cellules comprenant : un bras d'homologie gauche et droit, où soit le bras d'homologie gauche, soit le bras d'homologie droit code pour l'incorporation d'une édition génomique à un locus cible ; et une cassette à double sélection excisable située dans les bras d'homologie gauche et droit, où la cassette à double sélection excisable comprend : un premier marqueur de sélection ; et un second marqueur de sélection ; où le premier marqueur de sélection et le second marqueur de sélection se trouvent entre un premier et un second site d'excision. Des vecteurs de réparation dirigée par homologie (HDR) comprenant une construction selon la description, et des procédés d'utilisation desdits vecteurs sont en outre décrits.
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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: English (EN)
Filing Language: English (EN)