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Updated: Mar 18, 2026

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Practical Considerations for Using Pooled Lentiviral CRISPR Libraries.

Joel R McDade1, Nicole C Waxmonsky1, Lianna E Swanson1

  • 1Addgene, Cambridge, Massachusetts.

Current Protocols in Molecular Biology
|July 2, 2016
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 technology enables genome-wide screening by easily generating guide RNAs (gRNAs) for gene manipulation. This review details CRISPR/Cas9 basics, available libraries, and protocols for challenging screening experiments.

Keywords:
CRISPR/Cas9genome-wide screeningpooled libraries

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Area of Science:

  • Molecular Biology
  • Genetics
  • Bioengineering

Background:

  • CRISPR/Cas9 technology offers versatile gene editing capabilities, including knockout, repression, and activation.
  • The development of pooled lentiviral CRISPR libraries has increased accessibility for genetic research.

Purpose of the Study:

  • To provide an overview of CRISPR/Cas9 technology for genome-wide screening.
  • To describe publicly available CRISPR libraries.
  • To offer a general protocol for conducting CRISPR/Cas9-based genome-wide screening experiments.

Main Methods:

  • Review of CRISPR/Cas9 technology principles.
  • Compilation and description of existing pooled lentiviral CRISPR libraries.
  • Outline of a general protocol for genome-wide screening using CRISPR/Cas9.

Main Results:

  • CRISPR/Cas9 is well-suited for genome-wide screening due to ease of guide RNA generation and Cas9 versatility.
  • Despite widespread adoption for genetic experiments, genome-wide screening remains technically demanding.
  • Several pooled lentiviral CRISPR libraries are publicly available.

Conclusions:

  • CRISPR/Cas9 is a powerful tool for functional genomics and large-scale genetic screens.
  • Accessible CRISPR libraries and detailed protocols can facilitate complex genome-wide screening experiments.
  • Further optimization of protocols is needed to overcome technical challenges in CRISPR/Cas9 screening.