A comprehensive protocol resource for performing pooled shRNA and CRISPR screens

Leonie A. Cluse, Iva Nikolic, Deborah Knight, Piyush B. Madhamshettiwar, Jennii Luu, Karla J. Cowley, Timothy Semple, Gisela Mir Arnau, Jake Shortt, Ricky W. Johnstone, Kaylene J. Simpson

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)

Abstract

This chapter details a compendium of protocols that collectively enable the reader to perform a pooled shRNA and/or CRISPR screen—with methods to identify and validate positive controls and subsequent hits; establish a viral titer in the cell line of choice; create and screen libraries, sequence strategies, and bioinformatics resources to analyze outcomes. Collectively, this provides an overarching resource from the start to finish of a screening project, making this technology possible in all laboratories.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsJohn M. Walker
Place of PublicationNew York, NY
PublisherHumana Press
Chapter17
Pages201-227
Number of pages27
ISBN (Electronic)9781493975686
ISBN (Print)9781493975679
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
PublisherHumana Press, Inc.
Volume1725
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Drop out screen
  • Lentivirus
  • Pooled CRISPR
  • Pooled shRNA

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