	US 12,297,466 B2
	Engineered Cas9 nucleases
Barrett Ethan Steinberg, Somerville, MA (US); and Derek Cerchione, Cambridge, MA (US)
Assigned to Editas Medicine, Inc., Cambridge, MA (US)
Filed by Editas Medicine, Inc., Cambridge, MA (US)
Filed on Jul. 14, 2021, as Appl. No. 17/375,462.
Application 16/544,452 is a division of application No. 16/003,967, filed on Jun. 8, 2018, granted, now 10,428,319, issued on Oct. 1, 2019.
Application 17/375,462 is a continuation of application No. 16/544,452, filed on Aug. 19, 2019, granted, now 11,098,297.
Claims priority of provisional application 62/665,388, filed on May 1, 2018.
Claims priority of provisional application 62/517,811, filed on Jun. 9, 2017.
Prior Publication US 2022/0177861 A1, Jun. 9, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); C12N 1/21 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01)

CPC C12N 9/22 (2013.01) [C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)]

6 Claims

1. A method of editing a population of double stranded DNA (dsDNA) molecules, the method comprising contacting the dsDNA molecules with:

(i) a polypeptide comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 13, wherein the SPCas9-polypeptide comprises an amino acid substitution at two or more of the following positions of SEQ ID NO:13: D23, T67, Y128, and D1251, and

(ii) a guide nucleic acid having a region complementary to a target sequence of the dsDNA molecules,

whereby a plurality of the dsDNA molecules are edited.