US 11,939,604 B2
Modified cascade ribonucleoproteins and uses thereof
Stan Johan Jozef Brouns, Wageningen (NL); and John van der Oost, Renkum (NL)
Assigned to Caribou Biosciences, Inc., Berkeley, CA (US)
Filed by Caribou Biosciences, Inc., Berkeley, CA (US)
Filed on Feb. 18, 2021, as Appl. No. 17/179,215.
Application 17/179,215 is a continuation of application No. 16/914,203, filed on Jun. 26, 2020, granted, now 10,954,498.
Application 16/914,203 is a continuation of application No. 16/554,225, filed on Aug. 28, 2019, granted, now 10,711,257, issued on Jul. 14, 2020.
Application 16/554,225 is a continuation of application No. 15/802,413, filed on Nov. 2, 2017, granted, now 10,435,678, issued on Oct. 8, 2019.
Application 15/802,413 is a continuation of application No. 14/997,474, filed on Jan. 15, 2016, granted, now 9,885,026, issued on Feb. 6, 2018.
Application 14/997,474 is a continuation of application No. 14/326,099, filed on Jul. 8, 2014, abandoned.
Application 14/326,099 is a continuation of application No. 14/240,735, abandoned, previously published as PCT/EP2012/076674, filed on Dec. 21, 2012.
Claims priority of application No. 1122458 (GB), filed on Dec. 30, 2011.
Prior Publication US 2021/0171926 A1, Jun. 10, 2021
Int. Cl. C12N 9/22 (2006.01); A61K 48/00 (2006.01); C07K 14/245 (2006.01); C07K 14/47 (2006.01); C12N 9/16 (2006.01); C12N 15/62 (2006.01); C12N 15/66 (2006.01); C12N 15/70 (2006.01); C12N 15/74 (2006.01); C12N 15/81 (2006.01); C12N 15/82 (2006.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01); A61K 38/00 (2006.01)
CPC C12N 9/22 (2013.01) [A61K 48/005 (2013.01); C07K 14/245 (2013.01); C07K 14/47 (2013.01); C12N 9/16 (2013.01); C12N 15/62 (2013.01); C12N 15/66 (2013.01); C12N 15/70 (2013.01); C12N 15/74 (2013.01); C12N 15/81 (2013.01); C12N 15/82 (2013.01); C12N 15/86 (2013.01); C12N 15/902 (2013.01); C12N 15/907 (2013.01); A61K 38/00 (2013.01); C07K 2319/09 (2013.01); C07K 2319/22 (2013.01); C07K 2319/60 (2013.01); C07K 2319/71 (2013.01); C07K 2319/80 (2013.01); C07K 2319/85 (2013.01); C12N 2310/20 (2017.05); C12Y 301/21004 (2013.01)] 13 Claims
 
1. A method of modifying a target nucleic acid comprising:
contacting the target nucleic acid with a Type I CRISPR composition comprising:
a Type I CASCADE protein complex comprising a Cse1 subunit protein, having an N-terminus and a C-terminus,
a Cas3 K320N mutant protein, and
a CRISPR-derived RNA (crRNA) molecule comprising a spacer sequence complementary to the target nucleic acid;
wherein the Cas3 mutant protein comprises an Escherichia coli Cas3 mutant protein and the Cas3 mutant protein is fused to the N-terminus of the Cse1 subunit protein by a linker polypeptide.