US2022389460A1PendingUtilityA1

CRISPR-RELATED METHODS AND COMPOSITIONS WITH GOVERNING gRNAS

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Assignee: EDITAS MEDICINE INCPriority: Nov 7, 2013Filed: Jun 14, 2022Published: Dec 8, 2022
Est. expiryNov 7, 2033(~7.3 yrs left)· nominal 20-yr term from priority
Inventors:Feng ZhangDeborah PalestrantBeverly L. DavidsonJordi Mata-FinkEdgardo RodriguezAlexis Borisy
C12N 15/111C12N 9/22C12N 15/63C12N 2310/20C12N 15/1137C12N 2310/10C12N 2800/80C12N 15/11A61K 48/00C12N 15/902
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Claims

Abstract

Disclosed herein are methods and compositions useful in targeting a payload to, or editing a target nucleic acid, where a governing gRNA molecule is used to target, optionally inactivate, a Cas9 molecule or a Cas9 molecule/gRNA complex.

Claims

exact text as granted — not AI-modified
1 . A method of altering a eukaryotic cell or a target nucleic acid of a eukaryotic cell, the method comprising:
 contacting the eukaryotic cell or target nucleic acid of the eukaryotic cell with:   a) a nucleic acid encoding a first guide RNA (gRNA) molecule;   b) a Cas9 molecule, or a nucleic acid encoding a Cas9 molecule; and   c) a payload comprising an inhibitor of a transcription factor, a post-translational modification enzyme, or a post-transcriptional modification enzyme, or a drug that binds to the Cas9 molecule or the gRNA that activates or inhibits activity of the Cas9 molecule or the gRNA.   
     
     
         2 . The method of  claim 1 , wherein the inhibitor is a siRNA, a shRNA, a ribozyme, an antisense-oligonucleotide, an antibody, or an aptamer. 
     
     
         3 . The method of  claim 1 , wherein the inhibitor is a siRNA. 
     
     
         4 . The method of  claim 1 , wherein the inhibitor is a shRNA. 
     
     
         5 . The method of  claim 1 , wherein the inhibitor is a ribozyme. 
     
     
         6 . The method of  claim 1 , wherein the inhibitor is an antisense-oligonucleotide. 
     
     
         7 . The method of  claim 1 , wherein the inhibitor is an antibody. 
     
     
         8 . The method of  claim 1 , wherein the inhibitor is an aptamer. 
     
     
         9 . The method  claim 1 , wherein the drug is an entity that alters enzymatic activity, transcriptional activity, or translational activity of the Cas9 molecule or the gRNA. 
     
     
         10 . The method of  claim 1 , wherein the drug is an entity that alters the amount, distribution, or structure of the Cas9 molecule or the gRNA. 
     
     
         11 . The method of  claim 1 , wherein the drug is an entity that breaks a covalent or non-covalent bond in the Cas9 molecule or the gRNA. 
     
     
         12 . The method of  claim 1 , wherein the drug is an entity that attaches a moiety to the Cas9 molecule or the gRNA. 
     
     
         13 . The method of  claim 1 , wherein the drug is an entity that alters the three dimensional, secondary, tertiary, or quaternary structure, of the Cas9 molecule or the gRNA. 
     
     
         14 . The method of  claim 1 , wherein the drug is an entity that increases or decreases 3′ polyadenylation of the gRNA. 
     
     
         15 . The method of  claim 1 , wherein the drug is polyadenylate polymerase, cleavage and polyadenylation specificity factor (CPSF), or a poly(A) binding protein. 
     
     
         16 . The method of  claim 1 , wherein the drug is inhibitor of DNA binding 1 (ID1), inhibitor of DNA binding 2 (ID2), inhibitor of DNA binding 3 (ID3), or inhibitor of DNA binding 4 (ID4). 
     
     
         17 . The method of  claim 1 , wherein the Cas9 molecule is an  S. aureus  Cas9 molecule. 
     
     
         18 . The method of  claim 1 , wherein the Cas9 molecule is an  S. pyogenes  Cas9 molecule.

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