	US 12,344,838 B2
	CPF1 complexes with reduced indel activity
Feng Zhang, Cambridge, MA (US); Bernd Zetsche, Cambridge, MA (US); Winston Yan, Cambridge, MA (US); Neville Espi Sanjana, Cambridge, MA (US); and Sara Jones, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed by The Broad Institute, Inc., Cambridge, MA (US); and Massachusetts Institute of Technology, Cambridge, MA (US)
Filed on Feb. 7, 2022, as Appl. No. 17/666,248.
Application 17/666,248 is a continuation of application No. 16/095,253, granted, now 11,286,478, previously published as PCT/US2017/028461, filed on Apr. 19, 2017.
Claims priority of provisional application 62/324,834, filed on Apr. 19, 2016.
Prior Publication US 2022/0162584 A1, May 26, 2022
Int. Cl. C12N 15/10 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/63 (2006.01); C12N 15/90 (2006.01)

CPC C12N 15/102 (2013.01) [C12N 9/22 (2013.01); C12N 15/113 (2013.01); C12N 15/63 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12Y 305/04005 (2013.01)]

32 Claims

1. An engineered guide RNA (gRNA) capable of forming a CRISPR-Cpf1 complex with a Cpf1 effector protein and directing the CRISPR-Cpf1 complex to a target sequence; wherein the gRNA comprises a dead guide sequence capable of reducing indel activity of the CRISPR-Cpf1 complex; wherein

(a) said guide sequence is 10-14 nucleotides in length and complementary to the target sequence, or

(b)said guide sequence comprises a matching section and a mismatching section with the matching section being 10-16 contiguous nucleotides in length and complementary to the target sequence, wherein the mismatching section is present on the 3′ end of the gRNA.