RNAi Methods and Screening: RNAi Based High-Throughput Genetic Interaction Screening

Patricia G. Cipriani, Fabio Piano

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Expanding on decades of mutational analyses, numerous genome-scale RNAi screens have now been performed in C. elegans, leading to estimates that the majority of genes with essential functions that can be revealed by single-gene perturbations have already been identified in this organism. To build on this basic foundation and uncover condition-dependent or combinatorial effects of non-essential genes will require even higher-scale screening. Here we describe a method for performing high-throughput RNAi-based screens in C. elegans in liquid in 96-well plates, and we explain how to systematically test for enhancement and suppression of temperature-sensitive mutations. This chapter covers our entire set of protocols, from setting up the experiment and screening schedule, to scoring the results. The rapid acquisition of high-quality images of each experiment allows the management of a large number of samples per screening cycle and opens up new possibilities for quantitative scoring, computerized image analysis, and the ability to review results independent of the time constraints that are associated with large-scale screening.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
PublisherAcademic Press Inc.
Pages89-111
Number of pages23
DOIs
StatePublished - 2011

Publication series

NameMethods in Cell Biology
Volume106
ISSN (Print)0091-679X

Keywords

  • Amplification
  • Inoculation
  • Lethality
  • Lineage
  • Sensitivity
  • Synchronization

ASJC Scopus subject areas

  • Cell Biology

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  • Cite this

    Cipriani, P. G., & Piano, F. (2011). RNAi Methods and Screening: RNAi Based High-Throughput Genetic Interaction Screening. In Methods in Cell Biology (pp. 89-111). (Methods in Cell Biology; Vol. 106). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-544172-8.00004-9