US 12,264,331 B2
Systems and methods for one-shot guide RNA (ogRNA) targeting of endogenous and source DNA
Ari E. Friedland, Boston, MA (US); Hariharan Jayaram, Cambridge, MA (US); and Barrett Ethan Steinberg, Cambridge, MA (US)
Assigned to EDITAS MEDICINE, INC., Cambridge, MA (US)
Filed by EDITAS MEDICINE, INC., Cambridge, MA (US)
Filed on May 12, 2023, as Appl. No. 18/316,880.
Application 17/332,764 is a division of application No. 16/666,982, filed on Oct. 29, 2019, granted, now 11,028,411, issued on Jun. 8, 2021.
Application 16/666,982 is a division of application No. 15/980,450, filed on May 15, 2018, granted, now 10,494,649, issued on Dec. 3, 2019.
Application 15/980,450 is a division of application No. 15/934,750, filed on Mar. 23, 2018, granted, now 10,006,054, issued on Jun. 26, 2018.
Application 15/934,750 is a division of application No. 15/832,567, filed on Dec. 5, 2017, granted, now 9,963,719, issued on May 8, 2018.
Application 18/316,880 is a continuation of application No. 17/332,764, filed on May 27, 2021, granted, now 11,692,205.
Claims priority of provisional application 62/503,640, filed on May 9, 2017.
Claims priority of provisional application 62/430,154, filed on Dec. 5, 2016.
Prior Publication US 2023/0416787 A1, Dec. 28, 2023
Int. Cl. C12N 15/90 (2006.01); C12N 9/22 (2006.01); C12N 9/96 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/63 (2006.01)
CPC C12N 15/907 (2013.01) [C12N 9/22 (2013.01); C12N 9/96 (2013.01); C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 15/113 (2013.01); C12N 15/63 (2013.01); C12N 2310/20 (2017.05); C12N 2810/10 (2013.01)] 26 Claims
 
1. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a Cpf1 protein, wherein the nucleic acid sequence comprises a eukaryotic nucleic acid sequence that
a) is at least 17 nucleotides in length; and
b) comprises or is adjacent to a protospacer adjacent motif (PAM) that is recognized by the Cpf1 protein,
wherein the Cpf1 comprises an amino acid insertion, relative to SEQ ID NO: 13, at a position selected from the group consisting of:
i) between amino acid positions 147 and 148;
ii) anywhere between amino acid positions 484 and 492,
iii) anywhere between amino acid positions 568 and 590,
iv) anywhere between amino acid positions 795 and 855,
v) anywhere between amino acid positions 1131 and 1140, and
vi) anywhere between amino acid positions 1160 and 1173, and
where the insertion relative to SEQ ID NO: 13 results in a Cpf1 protein having at least 95% sequence identity to SEQ ID NO: 13.