Gene therapeutics for fibrodysplasia ossificans progressiva

Abstract

In some aspects, the disclosure relates to compositions and methods for treating fibrodysplasia ossificans progressiva (FOP) in a subject. In some aspects, the disclosure provides isolated nucleic acids, and vectors such as rAAV vectors, configured to express transgenes that inhibit (e.g., decrease) expression of mutated AVCR1 gene in muscle cells or connective tissues.

1 - 16 . (canceled) 
     
     
         17 . A recombinant gene editing complex comprising:
 (i) a single guide RNA (sgRNA) that specifically hybridizes to a target nucleic acid sequence of an ACVR1 gene;   (ii) a first rAAV particle encoding a first recombinant gene editing protein or fragment thereof; and   (iii) a second rAAV particle encoding a second recombinant gene editing protein or fragment thereof.   
     
     
         18 . The recombinant gene editing complex of  claim 17 , wherein the first recombinant gene editing protein comprises a Cas9-based adenine base editor (ABE) N-terminus portion and its fragments thereof and wherein the second recombinant gene editing protein comprises a Cas9-based adenine base editor (ABE) C-terminus portion and its fragments thereof, optionally wherein the second recombinant gene editing protein comprises a Cas9-based adenine base editor (ABE) C-terminus portion and its fragments thereof. 
     
     
         19 . The recombinant gene editing complex of  claim 17 , further comprising a protospacer adjacent motif. 
     
     
         20 . The recombinant gene editing complex of  17 , wherein the recombinant gene editing protein is a protein of the Crisper/Cas9 system. 
     
     
         21 . The recombinant gene editing complex of  17 , wherein the rAAV particles comprises an AAV9 capsid protein or variant thereof or an AAV6.2 capsid protein or variant thereof. 
     
     
         22 . The recombinant gene editing complex of  claim 18 , wherein each of the N-terminal ABE construct and the C-terminal ABE construct is divided by using a trans-splicing intein sequence. 
     
     
         23 . The recombinant gene editing complex of  claim 22 , wherein the N-terminal ABE construct comprises protospacer having the ACVR1-R206H mutation. 
     
     
         24 . The recombinant gene editing complex of  claim 22 , wherein the C-terminal ABE construct is integrated into the AAV6.2 or AAV9 capsid protein. 
     
     
         25 - 30 . (canceled)