Abstracts (complete list) - Wissenschaft Online

Carola Pongratz, Benjamin Yazdanapanah, Hamid Kashkar, Martin Kroenke
Antiviral implications of a convertible HIV-1 directed siRNA
library
RNA interference (RNAi) has been recognized as a powerful gene therapy strategy
against RNA viruses like human immunodeficiency virus (HIV). However, the efficacy of
transiently transfected or stably expressed RNAi is limited by the rapid development of
escape mutant viruses. Here we describe the construction of a genome-wide
randomized lentiviral RNAi library directed against HIV type-1. Employing appropriate
selection procedures we have selected approximately 484 short interfering RNA
sequences. Potent siRNA sequences were subcloned into a lentiviral expression cassette.
The possible regulatory effects of the shRNA vectors on HIV expression were assessed
by co-transfection with an HIV-derived pNL4.3-luc expression vector followed by a
luciferase assay. More than 50% of the lentiviral expression cassettes proved to inhibit
HIV-driven luciferase expression by greater than 70%. These siRNAs will be scrutinized
for their binding to conserved or variable regions of HIV, respectively, and tested for
their suitability of long-term inhibition of wild type HIV isolates.