Compositions and methods of nucleic acid-targeting nucleic acids

Abstract

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.