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Related Concept Videos

Genetic Screens02:46

Genetic Screens

5.1K
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
5.1K

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Updated: Oct 2, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Pooled CRISPR-Based Genetic Screens in Mammalian Cells

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Dissecting Molecular Phenotypes Through FACS-Based Pooled CRISPR Screens.

Oriana Genolet1, Liat Ravid Lustig1, Edda G Schulz2

  • 1Otto Warburg Laboratories, Max Planck Institute for Molecular Genetics, Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|February 26, 2022
PubMed
Summary

Pooled CRISPR screens enable researchers to study gene regulatory networks by analyzing how proteins respond to genetic changes. This method uses single-guide RNA (sgRNA) libraries and cell sorting to identify genes influencing specific cellular phenotypes.

Keywords:
CRISPR screenEmbryonic stem cellFlow-FISHIntracellular antibody stainingMAPK pathwayRNASignalingsgRNA library

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Last Updated: Oct 2, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
09:05

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

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Area of Science:

  • Molecular Biology
  • Genomics
  • Cell Biology

Background:

  • Pooled CRISPR screens are powerful for dissecting complex regulatory networks.
  • They assess protein responses to genetic perturbations in a multiplexed manner.
  • Single-guide RNA (sgRNA) per cell allows high-throughput genetic perturbation.

Purpose of the Study:

  • Provide detailed guidelines for designing and executing pooled CRISPR screens.
  • Focus on investigating molecular phenotypes.
  • Enable identification of genes regulating specific cellular behaviors.

Main Methods:

  • Generate custom sgRNA libraries.
  • Perform Fluorescence-Activated Cell Sorting (FACS) for phenotypic enrichment.
  • Utilize readouts like intracellular antibody staining or Flow-FISH for phosphorylation or RNA abundance analysis.

Main Results:

  • Demonstrate the application of pooled CRISPR screens for molecular phenotype investigation.
  • Showcase methods for custom sgRNA library generation and FACS-based screening.
  • Highlight the utility of various perturbation systems and readout options.

Conclusions:

  • Pooled CRISPR screens offer a versatile approach to study diverse molecular and cellular phenotypes.
  • The described guidelines facilitate the design and execution of such screens.
  • This technology expands the scope of research into gene function and regulation.