Development of conditional multi-shRNA constructs for anti-HIV-1 RNA-silencing

• Applicant: Jens Gruber, Ph.D.
• Subject Area: Biochemistry
• Term: from 2006 to 2009
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 26129913

The main aim of this project is to find new anti HIV-1 agents using RNA interference. To do this I will use conditional viral vectors to express short hairpin RNAs directed against multiple target sites along the HIV-1 RNA sequence. The shRNA expression will be directed against putative escape mutations as well as against genes that are essential for the virus. The antiviral activity of multimeric shRNAs under a single promoter will be compared to shRNAs under the control of individual polymerase III promoters to identify the optimal transcription conditions for efficient HIV-1 silencing. Subsequently combination siRNA-antiviral constructs will be tested in vitro, in peripheral blood cells and in T cell lines. Subsequent tests will be performed in vivo in a mouse model system. I will also ask whether HIV-1 encodes an RNAi suppressor that inhibits the host cell RNA silencing machinery. If anti- RNAi effects are observed in HIV-1 infected cells I will investigate whether this suppressor functions at the protein or RNA level. In this context I also plan to identify and catalogue HIV-1 encoded small RNA molecules, such as micro RNAs.

• DFG Programme: Research Fellowships
• International Connection: Netherlands
• Host: Professor Dr. Ben Berkhout