SG11201908596WA - Mmlv reverse transcriptase variants - Google Patents

Abstract

1 • 4 N QC O O r•i 00 O 1-1 r•i O (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 01 November 2018 (01.11.2018) WIPO I PCT ~ill IIIIIIIIIImillIVIIIIIIIHo o olo Iniolioo Imo oimIE (10) International Publication Number WO 2018/200867 Al (51) International Patent Classification: C12N 9/12 (2006.01) Cl 2N 15/33 (2006.01) C12N 9/22 (2006.01) (21) International Application Number: PCT/US2018/029641 (22) International Filing Date: 26 April 2018 (26.04.2018) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/490,492 26 April 2017 (26.04.2017) US (71) Applicant: 10X GENOMICS, INC. [US/US]; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). (72) Inventors: LEE, Josephine; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). MARRS, Sa- muel; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). MCDERMOTT, Geoffrey; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). MESCHI, Francesca; 7068 Koll Center Park- way, Suite 401, Pleasanton, California 94566 (US). MON- TESCLAROS, Luz; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94556 (US). PFEIFFER, Kather- ine; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). SHUGA, Joseph Francis; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). TERRY, Jessica Michele; 7068 Koll Cen- ter Parkway, Suite 401, Pleasanton, California 94556 (US). ZIRALDO, Solongo B.; 7068 Koll Center Parkway, Suite 401, Pleasanton, California 94566 (US). (74) Agent: SCHULTZ, Jason; WILSON SONSINI GOODRICH & ROSATI, 650 Page Mill Road, Palo Alto, US 94304 (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, (54) Title: MMLV REVERSE TRANSCRIPTASE VARIANTS 100 104 106 FIG. 1 (57) : Disclosed herein, are compositions, methods, and kits comprising engineered reverse transcription enzymes that ex- hibit several desired properties such as thermal stability, processive reverse transcription, non-templated base addition, and template switching ability. The engineered reverse transcription enzymes described herein demonstrate unexpectedly higher resistance to cell lysate inhibition, greater ability to capture full-length mRNA transcripts, and demonstrate improved results in small reaction volumes as compared to other engineered reverse transcription enzymes. [Continued on next page] WO 2018/200867 Al MIDEDIMOMOIDEIRDERI001010111MIIIMERVOIMIE 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, 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))