Behind the Scenes of RNA Interference 1

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Submitted by Timothy J. Kelly, Jesse R. Zamudio, and Phillip A. Sharp of the Koch Institute for Integrative Cancer Research

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RNA interference (RNAi) is a process by which small, targeted RNA molecules inhibit the expression of a gene. Functional RNAi requires an Argonaute (Ago) protein to bind a small RNA and together this complex targets specific RNAs for degradation. We have engineered a mouse embryonic stem cell line to express a single member of the Ago protein family (Ago2) and visualized its localization within cells alongside other cellular components. In this image, cellular cytoskeleton components (orange), DNA (blue) and a nuclear protein Cpsf6 (red) are visualized alongside Ago2 (green). We are utilizing this cell line to generate Ago2 interaction maps in normal cells in order to understand gene regulation in normal development and disease states. Introduction of small RNA molecules to the RNAi pathway in diseased cells may be employed in the next generation of cancer therapeutics to treat cancer in a specific, targeted manner. This technology holds the promise of therapeutics that exhibit reduced side effects as compared to many current options in cancer treatment.