CPC C12N 9/22 (2013.01) [C12N 15/11 (2013.01); C12N 15/86 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2740/15043 (2013.01)] | 38 Claims |
1. A variant of a reference guide ribonucleic acid (gRNA) (gRNA variant) capable of binding an engineered protein comprising a RuvC cleavage domain, wherein the RuvC cleavage domain comprises the sequence of amino acids 648-812 of SEQ ID NO: 2 with one or more amino acid modifications relative to the RuvC cleavage domain sequence, wherein:
a) the gRNA variant comprises at least one modification compared to the reference guide ribonucleic acid scaffold sequence, wherein the at least one modification is in a scaffold stem loop region, wherein the scaffold stem loop region comprises the sequence of SEQ ID NO: 245, or the sequence of SEQ ID NO: 245 with at least 1, 2, 3, 4, or 5 mismatches thereto; and wherein
b) the gRNA variant exhibits one or more improved characteristics compared to the reference guide ribonucleic acid.
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