Gene repair involving in vivo excision of targeting dna

Abstract

Methods of modifying, repairing, attenuating and inactivating a gene or other chromosomal DNA in a cell are disclosed. Also disclosed are methods of treating or prophylaxis of a genetic disease in an individual in need thereof.

1 . A method of treating or prophylaxis of a genetic disease in an individual in need thereof comprising the steps of:
 a) introducing into the individual cells which comprise a first vector comprising targeting DNA, wherein said targeting DNA is flanked by a restriction endonuclease site and comprises (1) DNA homologous to chromosomal DNA adjacent to a specific sequence of interest and (2) DNA which repairs the specific sequence of interest upon recombination between said targeting DNA and the chromosomal DNA; and   b) introducing into the individual a second vector comprising a nucleic acid encoding a restriction endonuclease which cleaves the restriction endonuclease site present in the first vector.   
     
     
         2 . The method of  claim 1  wherein the first vector is a viral vector. 
     
     
         3 . The method of  claim 2  wherein the second vector is a viral vector. 
     
     
         4 . The method of  claim 2  wherein the first vector is a plasmid. 
     
     
         5 . The method of  claim 1  wherein said targeting DNA is flanked by two restriction endonuclease sites. 
     
     
         6 . A method for treating or prophylaxis of a genetic disease in an individual in need thereof comprising the steps of:
 a) introducing into the individual cells which comprise a vector comprising targeting DNA, wherein said targeting DNA is flanked by a restriction endonuclease site and comprises (1) DNA homologous to chromosomal DNA adjacent to a specific sequence of interest and (2) DNA which repairs the specific sequence of interest upon recombination between said targeting DNA and the chromosomal DNA; and   b) introducing into the cell a restriction endonuclease which cleaves the restriction endonuclease site present in the vector.   
     
     
         7 . The method of  claim 6  wherein the vector is a viral vector. 
     
     
         8 . The method of  claim 6  wherein said targeting DNA is flanked by two restriction endonuclease sites.