Loss-of-Function RNAi Screen to Identify Necrosis-Signaling Molecules

  • David Mark Moquin
  • Francis Ka-Ming ChanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1857)


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.

Key words

Necroptosis Programmed necrosis Cell death Caspase-independent TNFα Transfection siRNA screen 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of ImmunologyDuke University Medical CenterDurhamUSA

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