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.
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