Loss-of-function RNAi screen to identify necrosis-signaling molecules

Abstract

Over the recent years, genome-wide RNA interference (RNAi) library screens have been instrumental in the identification of key regulators of various biological pathways. The prolific use of this technique is attributed to its amenability to a high-throughput format. Here, we present the step-by-step method to conduct a siRNA screen to identify genes involved in necroptosis, a nonapoptotic form of proinflammatory cell death. The method described here uses MTS cell proliferation assay to measure necroptosis, which is compatible with high-throughput format screening on multiwell microtiter plates. This ensures that the screen can be performed in a timely and efficient manner.