Sequences upstream of the CARP gene, vectors containing them and uses thereof

Abstract

The invention relates to novel promoter sequences derived from a portion upstream of the coding sequence of the gene for the CARP protein (Cardiac Ankyrin Repeat Protein), and which are capable of controlling the level and the specificity of expression of a transgene in vivo in cardiac muscle cells. The invention thus describes novel compositions, constructs, vectors and their uses in vivo for the transfer and expression of a nucleic acid in vivo in cardiac muscle cells. The subject of the present invention is also the use of the promoter sequences for generating transgenic animals which constitute models for studying certain cardiac pathologies.

1 - 39 . (canceled)  
     
     
         40 . A polynucleotide comprising a fragment of SEQ ID NO: 2 or a fragment having at least 90% sequence identity to a fragment of SEQ ID NO: 2, 
 wherein said polynucleotide in the absence of inverted terminal repeat sequences from human adeno-associated virus specifically induces expression in cardiac cells in vivo of a gene which is operably linked to said polynucleotide.    
     
     
         41 . An expression cassette comprising a sequence encoding a protein or an RNA of therapeutic interest operably linked to the polynucleotide according to claim  4 .  
     
     
         42 . The expression cassette according to  claim 41 , wherein the protein or RNA of therapeutic interest increases a rate of cardiac cell division, reduces or suppresses an immune response, induces angiogenesis, changes muscle contractility, reduces cardiac hypertrophy, reduces cardiac insufficiency, or reduces myocarditis.  
     
     
         43 . The expression cassette according to  claim 41 , wherein the protein or RNA of therapeutic interest is a vascular endothelial growth factor, a fibroblast growth factor, an angiopoietin, or a cytokine.  
     
     
         44 . The expression cassette according to  claim 41 , wherein the protein or RNA of therapeutic interest is an activating or an inhibiting transcription factor.  
     
     
         45 . The expression cassette according to  claim 41 , wherein the protein of therapeutic interest is an immunosuppressive protein.  
     
     
         46 . The expression cassette according to  claim 45 , wherein the immunosuppressive protein is interleukin-10, interleukin-2, or interleukin-8.  
     
     
         47 . The expression cassette according to  claim 41 , wherein the RNA of therapeutic interest is an antisense RNA or a ribozyme.  
     
     
         48 . The expression cassette according to  claim 41 , wherein the protein of therapeutic interest is nitric oxide synthetase, superoxide dismutase, or catalase.  
     
     
         49 . A vector comprising the polynucleotide according to  claim 40 .  
     
     
         50 . A vector comprising the expression cassette according to  claim 41 .  
     
     
         51 . The vector according to  claim 49 , further comprising an origin of replication which is active in cardiac cells.  
     
     
         52 . The vector according to  claim 49 , which is a plasmid or a cosmid.  
     
     
         53 . The vector according to  claim 49 , which is or is derived from an adenovirus, a retrovirus, a herpesvirus, or an adeno-associated virus.  
     
     
         54 . A composition comprising a therapeutically-effective amount of the polynucleotide according to  claim 40  and a pharmaceutically-acceptable carrier.  
     
     
         55 . A composition comprising a therapeutically-effective amount of the vector according to  claim 49  and a pharmaceutically-acceptable carrier.  
     
     
         56 . A transgenic nonhuman animal comprising a reporter gene operably linked to the polynucleotide according to  claim 40 .  
     
     
         57 . A method for expressing a protein or an RNA of therapeutic interest in cardiac cells in vivo, comprising 
 preparing a vector according to  claim 50 , and    introducing said vector into cardiac cells in vivo so that said protein or RNA of therapeutic interest is expressed.    
     
     
         58 . The vector according to  claim 49 , which is any DNA not encapsidated by viral proteins.