Human immunodeficiency virus-1 proviral gene disruption by targeted gene therapy: a hypothetical technique for the elimination of provirus from the infected cells

Abstract

A hypothetical technique is proposed for the elimination of all the integrated 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 protein would be constructed. Replication defective viral stocks would be made from this system which would have recombinant integrase-Rep78 protein packaged along with human immunodeficiency virus-1 long terminal repeat DNA. Since the Rep78 protein, which is a major regulatory protein of adeno-associated virus, has high affinity for human immunodeficiency virus-1 long terminal repeat, it would tether the newly synthesized 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 long terminal repeat would undergo homologous recombination with the provirus in the infected cells, facilitated by the nicking of the integrase part of the integrase-Rep78 recombinant protein. This selective gene knockout would be accomplished by the combined action of the chimeric 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, resulting in the replacement of human immunodeficiency virus-1 proviral genome with therapeutic DNA.