RNA interference strategies for localized gene silencing in cardiovascular tissues in vivo

• To address the functional role of a variety of molecules in vivo, we have developed RNA interference strategies for specific silencing of targeted genes. Since application of RNAi to in vivo gene silencing requires intracellular transcription and highly efficient transduction, we decided to employ a viral-mediated delivery system.
• We use adenoviral and AAV2 vectors to express U6 promoter-driven 21 base-pair hairpins targeted to a variety of gene sequences including Nox1, Nox2, Nox 4 and AT1a, along with enhanced green fluorescent protein (GFP) for monitoring transgene expression. Functionality, selectivity and specificity of the constructs are tested in vitro by transducing cells and analyzing target gene expression by real-time RT-PCR.
• These vectors are currently in use to silence specific Nox and other genes in select CNS regions, in cardiac progenitors and in blasocysts of BPH/5.