	US 12,110,490 B2
	CRISPR enzymes and systems
Feng Zhang, Cambridge, MA (US); Bernd Zetsche, Gloucester, MA (US); Fei Ran, Boston, MA (US); and James E. Dahlman, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US); and PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US)
Appl. No. 16/063,643
Filed by The Broad Institute, Inc., Cambridge, MA (US); Massachusetts Institute of Technology, Cambridge, MA (US); and President and Fellows of Harvard College, Cambridge, MA (US)
PCT Filed Dec. 16, 2016, PCT No. PCT/US2016/067201 § 371(c)(1), (2) Date Jun. 18, 2018, PCT Pub. No. WO2017/106657, PCT Pub. Date Jun. 22, 2017.
Claims priority of provisional application 62/324,811, filed on Apr. 19, 2016.
Claims priority of provisional application 62/269,860, filed on Dec. 18, 2015.
Prior Publication US 2019/0233814 A1, Aug. 1, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/00 (2006.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01)

CPC C12N 15/111 (2013.01) [C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/902 (2013.01); C12N 2310/20 (2017.05)]

33 Claims

1. A composition comprising:

a non-naturally occurring or engineered guide RNA (gRNA) comprising a guide sequence capable of hybridizing to a target sequence in a genomic locus of interest in a cell, wherein the gRNA is capable of forming a complex with a Cpf1 effector protein, wherein the gRNA comprises a loop comprising an aptamer sequence that is capable of binding to an adaptor protein; and

a mutated Cpf1 effector protein, wherein the mutated Cpf1 effector protein comprises at least one mutation, such that the mutated Cpf1 effector protein has no more than 5% of the nuclease activity of a corresponding wild-type Cpf1 effector protein not having the at least one mutation, and wherein the mutated Cpf1 effector protein comprises one or more nuclear localization sequences.