Lentiviral vector engineering for anti-HIV RNAi gene therapy.
RNA interference or RNAi-based gene therapy for the treatment of HIV-1 infection has recently emerged as a highly effective antiviral approach. The lentiviral vector system is a good candidate for the expression of antiviral short hairpin RNAs (shRNA) in HIV-susceptible cells. However, this strategy can give rise to vector problems because the anti-HIV shRNAs can also target the HIV-based lentiviral vector system. In addition, there may be self-targeting of the shRNA-encoding sequences within the vector RNA genome in the producer cell. The insertion of microRNA (miRNA) cassettes in the vector may introduce Drosha cleavage sites that will also result in the destruction of the vector genome during the production and/or the transduction process. Here, we describe possible solutions to these lentiviral-RNAi problems. We also describe a strategy for multiple shRNA expression to establish a combinatorial RNAi therapy.
Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam, University of Amsterdam, Amsterdam, The Netherlands.
Olivier
O
ter Brake
Jan-Tinus
JT
Westerink
Ben
B
Berkhout
D015316:Genetic Therapy; D005822:Genetic Vectors; D015658:HIV Infections / Q000628:therapy*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D034622:RNA Interference