231b Efficient Inhibition of HIV-1 Replication by RNA Interference Thijn R. Brummelkamp1, Atze T. Das2, Monique Vink2, René Bernards1, Ben Berkhout*2 1Dutch Cancer Institute, Amsterdam, the Netherlands and 2AMC, University of Amsterdam, the Netherlands
RNA interference provides a novel means for intracellular immunization of cells against HIV-1. Virus inhibition has recently been demonstrated in transient assays in which cells are treated with synthetic short interfering RNA (siRNA). We stably transduced a human T cell line with the pSUPER retroviral vector to express one of nine different siRNAs targeting the HIV-1 RNA genome. Profound cellular resistance against HIV-1 was observed exclusively for the siRNA that targets a sequence in the nef gene. Viral gene expression was reduced more than 10-fold by this siRNA-Nef. An escape virus was selected with a 106-basepair deletion in the non-essential nef gene which includes the target sequence. These results demonstrate the potential of siRNA-mediated gene therapy of HIV-susceptible cells, which could become a realistic strategy to overcome the devastating effect of HIV-1 on the immune system.
