COMPOSITIONS AND METHODS OF NUCLEIC ACID-TARGETING NUCLEIC ACIDS
This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.
1 - 26 . (canceled)
27 . A composition comprising:
a modified Cas9 protein, wherein the modified Cas9 protein comprises a modification of a region of a wild-type Cas9 protein selected from the group consisting of a Cas9 bridge helix region; a Cas9 highly basic patch within the Cas9 N-terminal region; a Cas9 HNH domain region; a Cas9 RuvC domain region; a Cas9 P-domain recognition region; an added enzymatic activity; insertion, deletion or substitution of a domain; insertion, deletion or substitution of a structural motif; a fusion; and combinations thereof.
28 . The composition of claim 27 , wherein the wild-type Cas9 protein is a Streptococcus pyogenes Cas9 protein.
29 . The compositions of claim 27 , wherein the modification is in the region of the wild-type Cas9 protein corresponding to amino acids 1-270 of a Streptococcus pyogenes Cas9.
30 . The compositions of claim 29 , wherein the modification is in the region corresponding to amino acids 50-100 of a Streptococcus pyogenes Cas9.
31 . The compositions of claim 30 , wherein the modification is in the region corresponding to amino acids 65-95 of a Streptococcus pyogenes Cas9.
32 . The compositions of claim 29 , wherein the modification is in the region corresponding to amino acids 22-49 of a Streptococcus pyogenes Cas9.
33 . The compositions of claim 27 , wherein the modification is in the region corresponding to amino acids 551-556 of a Streptococcus pyogenes Cas9.
34 . The compositions of claim 27 , wherein the modification is in the region corresponding to amino acids 445-507 of a Streptococcus pyogenes Cas9.
35 . The compositions of claim 27 , wherein the modification is in the region corresponding to amino acids 860-1100 of a Streptococcus pyogenes Cas9.
36 . The compositions of claim 27 , wherein the modification is in the region corresponding to amino acids 1096-1225 of a Streptococcus pyogenes Cas9.
37 . The compositions of claim 36 , wherein the modification is in the region corresponding to amino acids 1105-1138 of a Streptococcus pyogenes Cas9.
38 . The compositions of claim 27 , wherein the modification is in the region corresponding to amino acids 718-757 of a Streptococcus pyogenes Cas9.
39 . The composition of claim 27 , wherein the modification is an inserted nuclease domain or an inserted RNA binding domain.
40 . The composition of claim 39 , wherein the inserted RNA binding domain is Csy4.