Silencing of dux4 by recombinant gene editing complexes

Abstract

The disclosure relates to methods and compositions for regulating expression of DUX4. In some aspects, methods described by the disclosure are useful for treating a disease associated with aberrant DUX4 expression (e.g., facioscapulohumeral muscular dystrophy. FSHD).

1 - 20 . (canceled) 
     
     
         21 . A method for treating facioscapulohumeral muscular dystrophy (FSHD) in a subject in need thereof, the method comprising: administering to the subject a recombinant gene editing complex comprising:
 (i) a nucleic acid encoding a guide RNA (gRNA) that specifically hybridizes to a target nucleic acid sequence encoding DUX4; and   (ii) a CRISPR Cas protein fused to a transcriptional regulator domain, .   
     
     
         22 . The method of  claim 21 , wherein the subject is a a human. 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 21 to 23 , wherein the administration is by injection. 
     
     
         25 . The method of  claim 21 to 24 , wherein the recombinant gene editing complex is administered to muscle cells of the subject . 
     
     
         26 - 27 . (canceled) 
     
     
         28 . The method of  claim 21 , wherein the CRISPR Cas protein is a catalytically dead Cas9 protein (dCas9). 
     
     
         29 . The method of  claim 21 , wherein the CRISPR Cas protein is a Streptococcus pyogenes Cas protein (SpCas) or a Staphylococcus aureus Cas protein (SaCas). 
     
     
         30 . The method of  claim 21 , wherein the CRISPR Cas protein comprises the amino acid sequence set forth in SEQ ID NO: 42 or 45. 
     
     
         31 . The method of  claim 24 , wherein the administration is by intramuscular injection or intravenous injection. 
     
     
         32 . The method of  claim 21 , wherein the transcriptional regulator domain is a transcriptional repressor domain. 
     
     
         33 . The method of  claim 32 , wherein the transcriptional repressor domain comprises a Kruppel associated box domain (KRAB domain), a Chromo Shadow (CS) domain, or a WRPW domain. 
     
     
         34 . The method of  claim 32 , wherein the transcriptional repressor domain is a domain of an HP1α or Hes1 protein. 
     
     
         35 . The method of  claim 21 , wherein the transcriptional regulator domain is a transcriptional activator domain. 
     
     
         36 . The method of  claim 21 , wherein the gRNA specifically hybridizes to a target nucleic acid sequence encoding a DUX4 promoter. 
     
     
         37 . The method of  claim 21 , wherein the gRNA specifically hybridizes to a target nucleic acid sequence encoding exon 1 of DUX4. 
     
     
         38 . The method of  claim 21 , wherein the nucleic acid encoding the guide RNA (gRNA) comprises a U6 promoter operably linked to the gRNA. 
     
     
         39 . The method of  claim 21 , wherein the recombinant gene editing complex is administered to myoblasts, myocytes, or terminally differentiated muscle cells of the subject. 
     
     
         40 . A nucleic acid encoding:
 (a) a CRISPR Cas protein fused to a transcriptional regulator domain; and   (b) a guide RNA (gRNA) that specifically hybridizes to a target nucleic acid sequence encoding DUX4.   
     
     
         41 . The nucleic acid of  claim 40 , wherein the CRISPR Cas protein is a catalytically dead Cas9 protein (dCas9). 
     
     
         42 . The nucleic acid of  claim 40 , wherein the transcriptional regulator domain is a transcriptional repressor domain. 
     
     
         43 . The nucleic acid of  claim 40 , wherein the gRNA specifically hybridizes to a target nucleic acid sequence encoding a DUX4 promoter or exon 1 of DUX4.