Patent SG-11201903089R-A (SG)
Title: Aav delivery of nucleobase editors
Inventor: LIU DAVID
IPC: C12N5/10
Filed: 2017-10-13
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property 111111101111011101 HIM 0110101011111111111111H111111111011111111 Organization (10) International Publication Number International Bureau (43) International Publication Date .....0\"\" WO 2018/071868 Al 19 April 2018 (19.04.2018) WIP0 1 PCT (51) (21) (22) (25) Filing Language: (26) (30) (71) International Patent Classification: (72) Inventors: LIU, David, R.; 3 Whitman Circle, Lexington, C12N 7/00 (2006.01) A61K 48/00 (2006.01) MA 02420 (US). LEVY, Jonathan, Ma; 20 Sachem Street, C12N 15/864 (2006.01) C12N 15/10 (2006.01) Boston, MA 02120 (US). YEH, Wei, Hsi; 5 Cowperthwaite C12N 9/22 (2006.01) Cl 2N 15/79 (2006.01) Street, Cambridge, MA 02138 (US). C12N 5/10 (2006.01) (74) Agent: BAKER, C., Hunter; Wolf, Greenfield & Sacks, International Application Number: P.C., 600 Atlantic Avenue, Boston, MA 02210-2206 (US). PCT/US2017/056671 (81) Designated States (unless otherwise indicated, for every International Filing Date: kind of national protection available): AE, AG, AL, AM, 13 October 2017 (13.10.2017) 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, English DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, Publication Language: English HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, Priority Data: KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, 62/408,575 14 October 2016 (14.10.2016) US MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 62/475,780 23 March 2017 (23.03.2017) US 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, Applicant: PRESIDENT AND FELLOWS OF HAR- TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. VARD COLLEGE [US/US]; 17 Quincy Street, Cam- (84) Designated States (unless otherwise indicated, for every bridge, MA 02138 (US). kind of regional protection available): ARIPO (BW, GH, (54) —. respectively). Title: AAV DELIVERY OF NUCLEOBASE EDITORS 4.9 kilobases (AAV packaging limit) Intact base editor A-..... A N , ......N \ Spoas9 : . ir 1)/)/ I O N B 1 - /J G A r l i ll i R M10 Split base editor into a cell, e.g., via a recombinant The Cas9 protein or the nucleobase intein-C encoded by dnaEn and dnaEc, ligated together via intein-mediated or nucleobase editor, and methods i l i rg \ VA\ \ ' N'aiV \ \ \ Al'O N B SpCas9-N Inten-NEV AIN):91:1 \ , s, 4 • ,N .\ .... ± ' N rMITA \ - NV \ \ \.. ... A -..... A‘ ISpCas9-C /uGi ' Wet,61:1 using such AAV vectors and particles are also provided. Intein-C Figure 1A : Provided herein are methods of delivering \"split\" Cas9 protein or nucleobase editors virus (rAAV), to form a complete and functional Cas9 protein or nucleobase editor. is split into two sections, each fused with one part of an intein system (e.g., intein-N and Upon co-expression, the two sections of the Cas9 protein or nucleobase editor are splicing. Recombinant AAV vectors and particles for the delivery of the split Cas9 protein 1-1 adeno-associated ,-, GC pc (57) I N editor 0 GC protein 11 © of N O [Continued on next page] WO 2018/071868 Al MIDEDIM111101111111111111111111111101111111111111111111111011110111110111111111111 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). Published: — with international search report (Art. 21(3)) — before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) — with sequence listing part of description (Rule 5.2(a))