A Comprehensive Protocol Resource for Performing Pooled shRNA and CRISPR Screens

Methods Mol Biol. 2018:1725:201-227. doi: 10.1007/978-1-4939-7568-6_17.

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.

Keywords: Drop out screen; Lentivirus; Pooled CRISPR; Pooled shRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Computational Biology
  • Gene Library*
  • HEK293 Cells
  • High-Throughput Screening Assays*
  • Humans
  • Lentivirus / genetics*
  • RNA, Small Interfering / genetics*

Substances

  • RNA, Small Interfering