Patent SG-11201809552S-A (SG)
Title: Methods for breaking immunological tolerance using multiple guide rnas
Inventor: VORONINA VERA
IPC: C12N15/90
Filed: 2017-05-19
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property :::` , 111111H11101111110101110101110110101110101111111110111111011110111111 Organization International Bureau (10) International Publication Number i (43) International Publication Date .....•\"\" WO 2017/201476 Al 23 November 2017 (23.11.2017) WIPO I PCT (51) International Patent Classification: (72) Inventors: VORONINA, Vera; c/o Regeneron Pharma- A01K 67/027(2006.01) C12N 15/85 (2006.01) ceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, C07K 16/18 (2006.01) C07K 16/00 (2006.01) New York 10591 (US). MACDONALD, Lynn; c/o Re- C12N 15/90 (2006.01) generon Pharmaceuticals, Inc., 777 Old Saw Mill River (21) International Application Number: Road, Tarrytown, New York 10591 (US). PRISSETTE, PCT/US2017/033648 Marine; c/o Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, New York 10591 (US). LAI, (22) International Filing Date: 19 May 2017 (19.05.2017) Ka-Man Venus; 127 West Main Street, Unit 201, Tar- rytown, New York 10591 (US). BADITHE, Ashok; c/o Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill Riv- (25) Filing Language: English er Road, Tarrytown, New York 10591 (US). MURPHY, (26) Publication Language: English Andrew J.; c/o Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, New York 10591 (US). (30) Priority Data: DROGUETT, Gustavo; c/o Regeneron Pharmaceuticals, 62/339,472 20 May 2016 (20.05.2016) US Inc., 777 Old Saw Mill River Road, Tarrytown, New York 62/368,604 29 July 2016 (29.07.2016) US 10591 (US). FRENDEWEY, David; c/o Regeneron Phar- (71) Applicant: REGENERON PHARMACEUTICALS, maceuticals, Inc., 777 Old Saw Mill River Road, Tarry- INC. [US/US]; 777 Old Saw Mill River Road, Tarrytown, town, New York 10591 (US). ZAMBROWICZ, Brian; c/ New York 10591-6707 (US). o Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, New York 10591 (US). (54) Title: METHODS FOR BREAKING IMMUNOLOGICAL TOLERANCE USING MULTIPLE GUIDE RNAS sgRNA sgRNA -P 15T.:7•Ziqt-M4-6j Target 1 . Design sgRNA Combinations Single Targeting to Generate 2. Generate soRNAs 3. Eiectrcoorato V1-3 or ULC ES Celis Homozygous 4. TAQMAN® Screen for Biallelic Deletion Collapse 5. Microinjection 6. Screen FO Animals for Bialielic Deletion Homozygous Collapse FO - Homozygous KO 4-5 Months 1-1 FIG. 3 IN 71 , 1-1 (57) : Methods and compositions are provided for making non-human animals with reduced tolerance of a foreign antigen of 0 interest and making antigen-binding proteins against that foreign antigen of interest using such animals. The methods and compositions C•1 ----. employ CRISPR/Cas9 systems using multiple guide RNAs to reduce or eliminate expression of a self-antigen homologous to or sharing IN an epitope of interest with the foreign antigen of interest or to reduce or eliminate expression of an epitope on the self-antigen that is C:::;) shared with the foreign antigen of interest. C•1 C [Continued on next page] WO 2017/201476 Al IMEDIMOM0101011M0101110010101ROEHOHROMOVOIMIE (74) Agent: SCHILLING, Stephen H. et al.; Alston & Bird LLP, 101 South Tryon Street, Bank of America Plaza, Suite 4000, Charlotte, North Carolina 28280-4000 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): 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, 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. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (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), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: — as to the identity of the inventor (Rule 4.17(0) — as to the applicant's entitlement to claim the priority of the earlier application (Rule 4.17(iii)) Published: — with international search report (Art. 21(3)) — with sequence listing part of description (Rule 5.2(a))