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In Vivo CRISPR/Cas9 Screening to Simultaneously Evaluate Gene Function in Mouse Skin and Oral Cavity

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Genome-scale CRISPR screening in a single mouse liver.

Heather R Keys^1,2, Kristin A Knouse^3,2,4

¹Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.

|January 16, 2023

Summary

This study introduces a new method for high-throughput functional genomics in live mice, enabling genetic dissection of complex biological processes. The approach successfully identified novel pathways regulating liver cell fitness, advancing in vivo genetic studies.

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Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes

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Published on: May 23, 2025

Area of Science:

Genetics
Genomics
Mammalian Physiology

Background:

Understanding genetic determinants of mammalian physiology and disease is limited by current genetic dissection capacities.
Genome-wide CRISPR screening is effective for cellular processes but largely restricted to ex vivo systems due to delivery challenges.
There is a need for accessible high-throughput functional genomics in vivo.

Purpose of the Study:

To establish a genome-wide screening method for functional genomics in the liver of a single mouse.
To uncover genetic regulation of hepatocyte fitness using this novel in vivo approach.
To demonstrate the utility of in vivo genetic dissection for discovering biological pathways.

Main Methods:

Development of an accessible and scalable genome-wide CRISPR screening approach.
Application of the screening method to the liver of a single living mouse.
Uncovering genetic regulation of hepatocyte fitness.

Main Results:

Successfully established genome-wide screening in the liver of a single mouse.
Identified novel genetic pathways regulating hepatocyte fitness.
Discovered pathways not previously identified in cell culture screens, highlighting the value of in vivo studies.

Conclusions:

The developed approach provides a foundation for high-throughput functional genomics in living mammals.
This method is accessible, scalable, and adaptable for diverse phenotypes and applications.
Enables comprehensive investigation of mammalian physiology and disease through in vivo genetic dissection.