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Related Concept Videos

Genetic Screens02:46

Genetic Screens

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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
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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Related Experiment Video

Updated: Jan 7, 2026

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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A next-generation dual guide CRISPR system for genetic interaction library screening.

Thomas Burgold1, Emre Karakoc1,2, Emanuel Gonçalves1,3,4

  • 1Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.

Nature Communications
|December 6, 2025
PubMed
Summary
This summary is machine-generated.

We developed a new CRISPR/Cas9 system for efficient screening of gene interactions. This method simplifies cloning and improves dual guide RNA expression, enabling large-scale genetic interaction studies for cancer therapy development.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 enables gene function perturbation for genetic interaction screening.
  • Existing dual guide RNA systems face challenges in cloning, guide pair purity, and expression balance.

Purpose of the Study:

  • To develop an improved system for dual guide RNA delivery for large-scale CRISPR/Cas9 genetic screens.
  • To overcome limitations of current dual guide expression systems.

Main Methods:

  • A novel dual guide RNA delivery system utilizing tRNA spacers for single-step cloning.
  • Implementation with Streptococcus pyogenes Cas9 (SpCas9) for library-scale screening.
  • Screening of a 100,136 guide pair library in colorectal cancer cells.

Main Results:

  • The new system allows for a single-step cloning strategy with minimal undesired guide pairs (as low as 2%).
  • Achieved highly balanced expression of the two delivered guide RNAs.
  • Successfully identified synthetic lethal genetic interactions in colorectal cancer cells.

Conclusions:

  • This next-generation system facilitates efficient and large-scale genetic interaction screening.
  • The versatile system can be adapted for various CRISPR applications, including combination therapy identification.
  • Demonstrates a robust method for discovering gene interactions relevant to cancer research.