Patent SG-11201901364V-A (SG)
Title: Engineered target specific nucleases
Inventor: MILLER JEFFREY C
IPC: C12N9/22
Filed: 2017-08-24
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 01 March 2018 (01.03.2018) WIPO I PCT °mons °nolo ollloolmo Ho mil ioluom oimIE (10) International Publication Number WO 2018/039448 Al IGIHI (51) International Patent Classification: C12N 9/14 (2006.01) A61K 38/16 (2006.01) C12N 9/22 (2006.01) (21) International Application Number: PCT/US2017/048409 (22) International Filing Date: 24 August 2017 (24.08.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/378,978 24 August 2016 (24.08.2016) US 62/443,981 09 January 2017 (09.01.2017) US (71) Applicant: SANGAMO THERAPEUTICS, INC. [US/US]; 501 Canal Blvd., Suite A100, Richmond, Califor- nia 94804 (US). (72) Inventors: MILLER, Jeffrey C.; c/o Sangamo Therapeu- tics, Inc., 501 Canal Blvd., Suite A100, Richmond, Califor- nia 94804 (US). REBAR, Edward J.; c/o Sangamo Thera- peutics, Inc., 501 Canal Blvd., Suite A100, Richmond, Cal- ifornia 94804 (US). (74) Agent: PASTERNAK, Dahna; PASTERNAK PATENT LAW, 1900 Embarcadero Rd., Suite 211, Palo Alto, Cali- fornia 94303 (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, 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. (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, (54) Title: ENGINEERED TARGET SPECIFIC NUCLEASES 416 418 QL VEISELEEMMSELMIIKLMYVPHEYIELIEIMAIMIMIM CAGCTGGTGAAGAGCGAGCTGGAGGAGAAGAAGTCOGAGCTGCGOCACAAGCTGAAGTACGTGCCCCACGAGTACATCGAGCTGATCGAGATCGCCAGGAACAGC 422 448 450 170711711711711LIEIMIKI V M E F F MMV Y G YEIGIKIHIL SIGISIRIKIP OG A I ACCCAGGACCGCATCOTGGAGATGAAGGTGATGGAGTTOTTCATSAAGGTGTACGGCTACAGGGGARAGCACCTGGGCGGAAGCAGMAGCCTGACGGCGCCATC 467 469 476 479, 480, 481 486 Y T V GS P ID Y G V I V OMOIAIYIISIGI 4ElY TATACAGTGGGCAGGCCCATCGAT2ACGGCMGATCGTGGACACAAAGGCCTACAGCGGCGSCTACAATCTGCCTATCGGCCAGGCCGACGAGATGTAGAGATAC 490 496 499 T E EINIQI TMNMH I N I P I N I E tic WIM V Y P S S V T E FMF L F GTGGTGGAGAACCP,GACCCGGAITAP,GOACAT2CAACCCCAACGAGTGGTSGAAGGTGTACCCTAGCAGCGTGACCGAGT2CAAGTPCOTGT2CGTGACGGCCAC 525 527 531 537, 538 F EINIYIKIEML TEILMH I TMCMG A V L S V EEL L I G G EM I TTCAAGGGCAACTACAAGGCCCAGCTGACCAGGCTGAACACF2CACCAACTGCAATGGCGCCGTGCTGAGCGTGGAGGAGCTGCTGATCGGCGGCGAGATGATC MA GT L TL EE VMIRIKIFMMG E IMF (196 RESIDUE) AAAGCCGGOACCCTGACACTGGAGGAGGTGCGSCGCAAGTTCAACAACGGCGAGATCAACTTC OH BASES) POSITIONS MUTATED AND CHAREACTERIZED FOR IMPROVEMENTS IN ACTIVITY AND/OR SPECIFICITY 1-1 0 ACTIVE SITE RESIDUE RESIDUE MUTATED IN ELD/KKR VARIANTS 71' RESIDUE NUMBERS ATE INDICATED FOR ACTIVE SITE RESIDUES, ELD/KKR VARIANTS, AND HEAVILY MUTATED POSITIONS 71' 01 FIG. 1 O 00 (57) : Described herein are engineered nucleases comprising mutations in the cleavage domain (e.g., Fold or homologue 0 thereof) and/or DNA binding domain (zinc fmger protein, TALE, single guide RNA) such that on-target specificity is increased. o [Continued on next page] WO 2018/039448 Al MIDEDIMOMMIDIIMERINIMIIMMOHOHIMMEEM 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))