Use of meganucleases for inducing homologous recombination ex vivo and in toto in vertebrate somatic tissues and application thereof

Abstract

Use of meganucleases for inducing homologous recombination ex vivo and in toto in vertebrate somatic tissues and to its application for genome engineering and gene therapy.

1 - 18 . (canceled)  
     
     
         19 . Use of at least one meganuclease for the preparation of a medicament for preventing, improving or curing a genetic disease in the liver of a vertebrate in need thereof, said medicament being administered by any means to said vertebrate.  
     
     
         20 . Use of at least one meganuclease for the preparation of a medicament for preventing, improving or curing a disease caused by an infectious agent that presents a DNA intermediate, in the liver of a vertebrate in need thereof, said medicament being administered by any means to said vertebrate.  
     
     
         21 . Use of at least one meganuclease for genome engineering of non-human vertebrate liver tissue for non-therapeutic purpose, by introducing said meganuclease into the body of said non-human vertebrate.  
     
     
         22 . The use of  claim 19 , characterized in that said meganuclease can be used either as a polypeptide or as a polynucleotide construct comprising the sequence encoding said polypeptide under the control of appropriate transcription regulatory elements including a promoter, for example a tissue specific and/or inducible promoter.  
     
     
         23 . The use of  claim 19 , characterized in that said meganuclease is administered alone or in association with either at least an appropriate vehicle or carrier and/or with a targeting DNA.  
     
     
         24 . The use of  claim 19 , characterized in that said appropriate vehicle is selected in the group consisting of liposomes, polyethyleneimine and membrane translocating peptides.  
     
     
         25 . The use of  claim 19 , characterized in that said targeting fragment of DNA comprises a sequence which modifies the site of interest flanked by sequences sharing homologies to a targeted locus.  
     
     
         26 . The use of  claim 25 , characterized in that said homologous sequences have at least 50 bp, preferably more than 100 bp and more preferably more than 200 bp with said targeted locus.  
     
     
         27 . The use of  claim 25 , characterized in that said sequence which modifies the site of interest is the correct sequence of a gene for repairing a genetic lesion.  
     
     
         28 . The use of  claim 19 , characterized in that said meganuclease is selected from the group consisting of a homing endonuclease, a zinc-finger nuclease or a meganuclease variant derived from any initial meganuclease, said variant presenting a recognition and cleavage site different from the site of the initial one.  
     
     
         29 . The use of  claim 19 , characterized in that said sequence encoding said meganuclease and/or said targeting fragment of DNA is inserted in a vector.  
     
     
         30 . The use of  claim 19 , characterized in that said meganuclease is substantially non-immunogenic.  
     
     
         31 . The use of  claim 19 , characterized in that said meganuclease is used in association with a targeting fragment of DNA as defined in claim  7 , comprising a sequence able to repair the site of interest, for preventing, improving or curing a monogenetic recessive disease.  
     
     
         32 . The use of  claim 19 , characterized in that said meganuclease is used alone or in association with at least one appropriate vehicle and/or carrier for preventing, improving or curing genetic diseases caused by dominant or compound heterozygous mutations.  
     
     
         33 . The use of  claim 20 , characterized in that said infectious agent is a virus.  
     
     
         34 . The use of  claim 33 , characterized in that said virus is HBV.  
     
     
         35 . The use of  claim 19 , characterized in that said sequence encoding said meganuclease and said targeting fragment of DNA are inserted in the same vector.