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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
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
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Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM.

Alan S L Wong1, Gigi C G Choi1, Cheryl H Cui2

  • 1Synthetic Biology Group, Massachusetts Institute of Technology (MIT) Synthetic Biology Center, MIT, Cambridge, MA 02139; Research Laboratory of Electronics, MIT, Cambridge, MA 02139;

Proceedings of the National Academy of Sciences of the United States of America
|February 12, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces CombiGEM-CRISPR, a platform for screening gene combinations to find effective drug pairs. It accelerates the discovery of genetic and drug combinations for human diseases.

Keywords:
CRISPR-CasCombiGEMgenetic perturbationshigh-throughput screeningmultifactorial genetics

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Area of Science:

  • Genomics
  • Cancer Biology
  • Drug Discovery

Background:

  • Discovering gene and drug combinations to control cell phenotypes is challenging and difficult to scale.
  • Systematic identification of genetic interactions is crucial for understanding complex biological processes and developing targeted therapies.

Purpose of the Study:

  • To develop a scalable platform for massively parallel screening of combinatorial gene perturbations in human cells.
  • To translate identified gene pairs into effective drug combinations for modulating human disease phenotypes, specifically ovarian cancer.

Main Methods:

  • Utilized CRISPR-Cas9 and Combinatorial Genetics en masse (CombiGEM) to create barcoded combinatorial genetic libraries.
  • Performed high-throughput pooled screening of 23,409 dual guide-RNA combinations to identify gene pairs inhibiting ovarian cancer cell growth.
  • Validated gene pair effects using CRISPR-Cas knockouts and RNA-interference, and tested synergistic drug pairs.

Main Results:

  • Identified specific gene pairs, including epigenetic regulators KDM4C/BRD4 and KDM6B/BRD4, that inhibit ovarian cancer cell growth.
  • Demonstrated that drug pairs targeting identified gene pairs exhibit synergistic antiproliferative effects.
  • Successfully validated the CombiGEM-CRISPR platform's ability to identify impactful genetic and drug combinations.

Conclusions:

  • The CombiGEM-CRISPR platform offers a powerful tool for accelerating the systematic discovery of gene and drug combinations.
  • This technology has broad applicability for various biological settings and can advance the development of novel therapeutic strategies for complex diseases.
  • The study highlights the potential for translating genetic discoveries into effective combination therapies.