HUMAN IMMUNODEFICIENCY VIRUS-1 PROVIRAL GENE DISRUPTION BY TARGETED GENE-THERAPY - A HYPOTHETICAL TECHNIQUE FOR THE ELIMINATION OF PROVIRUSFROM THE INFECTED-CELLS

A hypothetical technique is proposed for the elimination of all the in tegrated human immunodeficiency virus-1 provirus from infected cells, based on the developing technology of selective gene excision through homologous recombination. In this technique, a recombinant retroviral packaging cell-line which would produce integrase-Rep78 chimeric prote in would be constructed. Replication defective viral stocks would be m ade from this system which would have recombinant integrase-Rep78 prot ein packaged along with human immunodeficiency virus-1 long terminal r epeat DNA. Since the Rep78 protein, which is a major regulatory protei n of adeno-associated virus, has high affinity for human immunodeficie ncy virus-1 long terminal repeat, it would tether the newly synthesize d human immunodeficiency virus-1 long terminal repeat (therapeutic DNA ) to the human immunodeficiency virus-1 proviral site in the infected cell. This newly reverse transcribed human immunodeficiency virus-1 lo ng terminal repeat would undergo homologous recombination with the pro virus in the infected cells, facilitated by the nicking of the integra se part of the integrase-Rep78 recombinant protein. This selective gen e knockout would be accomplished by the combined action of the chimeri c integrase Rep78 protein, where the Rep78 part would help docking of the therapeutic DNA to the proviral integration site and the integrase would provide nicking activity after homologous recombination, result ing in the replacement of human immunodeficiency virus-1 proviral geno me with therapeutic DNA.