Crispr-cas effector polypeptides and methods of use thereof
AssigneeThe Regents of the University of California
InventorsJennifer A. Doudna, Evangelina Nogales De La Morena, Jun-Jie Liu, Connor Andrew Tsuchida
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Abstract
The present disclosure provides variant CRISPR-Cas effector polypeptides, as well as engineered guide nucleic acids, and systems comprising the same. The present disclosure provides methods of modifying a target nucleic acid, using a variant CRISPR-Cas effector polypeptide of the present disclosure.
Claims66
A variant CRISPR-Cas effector polypeptide comprising an amino acid sequence having at least 50% amino acid sequence identity to the amino acid sequence depicted in FIG. 1A or FIG. 2A, wherein the variant CRISPR-Cas effector polypeptide comprises:
- i) one or more amino acid substitutions and/or insertions in the helical II domain relative to a reference CRISPR-Cas effector polypeptide comprising the amino acid sequence depicted in FIG. 1B or FIG. 2B;
- and/or
- ii) one or more amino acid substitutions and/or insertions in the RuvC domain relative to a reference CRISPR-Cas effector polypeptide comprising the amino acid sequence depicted in FIG. 1B or FIG. 2B, wherein said one or more amino acid substitutions and/or insertions provides for increased binding and/or cleavage of a target nucleic acid compared to the binding and/or cleavage of the target nucleic acid by the reference CRISPR-Cas effector polypeptide.
The variant CRISPR-Cas effector polypeptide of 6, wherein the variant CRISPR-Cas effector polypeptide comprises an amino acid sequence having at least 50% amino acid sequence identity to the amino acid sequence depicted in FIG. 2A, and wherein the helical II domain comprises the amino acid sequence HKKREGSLENP (SEQ ID NO:35) or an amino acid sequence having from 1 to 5 amino acid substitutions compare to HKKREGSLENP (SEQ ID NO:35).
Dependencies are inferred from the claim text.