Pooled shRNA Screening in Mammalian Cells as a Functional Genomic Discovery Platform

Methods Mol Biol. 2016:1470:49-73. doi: 10.1007/978-1-4939-6337-9_5.

Abstract

Functional genomic screens using shRNA technology are a great tool in biomedical research. As more labs gain access to the necessary reagents and technology to perform such screens, some may lack in-depth knowledge on the difficulties often encountered. With this protocol, we aim to point out the most important caveats of performing shRNA based screens and provide a streamlined workflow that can be easily adapted to meet the specific needs of any particular screening project.

Keywords: Functional genetics; RNAi; Screens; shRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Gene Library
  • Genomics / methods*
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Lentivirus / genetics
  • Mammals / genetics
  • Polymerase Chain Reaction / methods
  • RNA, Small Interfering / analysis
  • RNA, Small Interfering / genetics*
  • Transduction, Genetic
  • Workflow

Substances

  • RNA, Small Interfering