Genome-wide rnai screens for RNA processing events in drosophila melanogaster s2 cells

Natoya Peart, Eric Wagner

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Over the past 10 years, the design and application of genome-wide screening (GWS) has improved to the point that it can now be done at level of the individual laboratory. The advantages of GWSs compared to classical genetic screens include: immediate identification of a positive scoring gene, relatively short period of time necessary to conduct the screen (as little as 1 week), cell lines do not present developmental needs for gene expression that an organism normally would, and validation/confirmation of results is straightforward. Here, we describe a general protocol for GWS to be conducted in Drosophila melanogaster S2 cells. We provide specific details on what type of experiments must be done before initiating a screen, the materials that are required to conduct a screen, and make suggestions on methods to carry out secondary screening and counter-screening once the initial GWS is complete. Multiple considerations are also raised that focus on how to anticipate false positives/negatives and how to minimize their occurrence through intelligent design. Finally, we provide specific examples of data that our group has gathered from published genome-wide screens in order to exemplify how “hits” are scored and confirmed.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages235-245
Number of pages11
Volume1648
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1648
ISSN (Print)1064-3745

Keywords

  • Drosophila melanogaster
  • Gain-of-function
  • High-throughput
  • Loss-of-function
  • Reporter-based
  • RNAi

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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