Abstract

RNA interference (RNAi) allows for transient, targeted depletion of cellular or viral proteins. Previously, small interfering RNA (siRNA) screens targeting cellular factors successfully identified several host genes that are required for VACV infection, and other viruses such as HIV. In this chapter, we outline how RNAi can be adapted to unravel the functions of poxvirus genes, using a 96-well format. Additionally, we describe two different high-throughput methods (flow cytometry and automated microscopy) to assess infection levels of an engineered VACV that encodes a fluorescent reporter protein under an early and/or late viral gene promoter.

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