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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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Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish
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Scalable CRISPR Screens in Zebrafish Using MIC-Drop.

Saba Parvez1, Tejia Zhang1, Randall T Peterson2

  • 1Department of Pharmacology & Toxicology, University of Utah, Salt Lake City, UT, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 5, 2023
PubMed
Summary

Multiplexed Intermixed CRISPR Droplets (MIC-Drop) enables large-scale in vivo genetic screens in zebrafish. This novel platform overcomes previous challenges, making gene function studies more accessible and efficient.

Keywords:
CRISPR-Cas9F0 CRISPR screenGenetic screenHigh-throughputIn vivoReverse geneticsZebrafish

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Area of Science:

  • Genetics
  • Molecular Biology
  • Zebrafish Models

Background:

  • CRISPR-Cas9 is a key technology for gene function studies.
  • Large-scale in vivo genetic screens are difficult due to cost and labor.
  • Previous methods limit systematic gene interrogation in whole organisms.

Purpose of the Study:

  • To present a novel platform for large-scale reverse genetic screens in zebrafish.
  • To detail a protocol for implementing Multiplexed Intermixed CRISPR Droplets (MIC-Drop).
  • To facilitate efficient in vivo gene function analysis.

Main Methods:

  • Development of the MIC-Drop platform.
  • Utilizing CRISPR-Cas9 for multiplexed genetic perturbations.
  • Application in zebrafish for large-scale screening.

Main Results:

  • MIC-Drop enables scalable in vivo genetic screens.
  • The platform simplifies the process of generating and tracking mutant animals.
  • Facilitates systematic interrogation of gene function in a whole organism.

Conclusions:

  • MIC-Drop significantly advances the feasibility of large-scale reverse genetic screens in zebrafish.
  • This technology reduces barriers to in vivo genetic studies.
  • Enables more comprehensive understanding of gene function in a complex biological system.