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US 12,312,613 B2Granted
Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants
AssigneeThe General Hospital Corporation
InventorsBenjamin Kleinstiver, Russell T. Walton
Life of this patent
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Abstract
Streptococcus pyogenes Cas9 (SpCas9) variants with relaxed PAM requirements capable of high-resolution editing for various applications, and methods of use thereof.
Claims38
An isolated Streptococcus pyogenes Cas9 (SpCas9) protein, comprising an amino acid sequence that has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:1, wherein the amino acids at positions:
- 1135, 1136, 1218, 1219, 1335, and 1337 are:
- LWKQQR (“SpG”);
- LWRQQR;
- LWSQQR;
- LWKHQR;
- LWKSQR;
- LWRSQR;
- LWRSQK;
- LWSHQR;
- LWRHQR;
- LWRQQK;
- LWSQQK;
- LSKQQR;
- LWKQQK;
- LSKHQR;
- LWKSQK;
- LSRHQR;
- LWRVQK;
- LFRQQR;
- LSRQQR;
- LSRSQR;
- LARQQR;
- LSRVQR;
- ASREQR;
- WSREQR;
- LSREQR;
- FSREQR;
- LSKSQR;
- LWKVQK;
- LWKHQK;
- LWSSQK;
- LWSHQK;
- LWSSQR;
- LWRVQR;
- LSKVQR;
- LWRHQK;
- LSSQQR;
- LWKVQR;
- LWSVQK;
- LSSHQR;
- LWSVQR;
- LSSVQR;
- LSKQQK;
- LSRVQK;
- LSKVQK;
- LSSSQR;
- LSKSQK;
- LSSVQK;
- LSRQQK;
- LSSQQK;
- LSRSQK;
- or LSKHQK, respectively.
49 more elements
Dependencies are inferred from the claim text.