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In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

Genetic Screens

Genetic Screens

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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Related Experiment Video

Updated: May 1, 2026

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Published on: February 17, 2016

A conditional knockout resource for the genome-wide study of mouse gene function.

William C Skarnes¹, Barry Rosen, Anthony P West

¹Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK. skarnes@sanger.ac.uk

|June 17, 2011

Summary

Researchers developed a high-throughput pipeline for creating reporter-tagged, conditional gene alleles in mouse embryonic stem cells. This scalable method accelerates the generation of targeted mutations for understanding gene function across mammals.

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Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

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Area of Science:

Genomics and Molecular Biology
Mammalian Genetics
Stem Cell Biology

Background:

Gene targeting in embryonic stem cells is crucial for mouse genome manipulation and conditional mutagenesis.
Large-scale knockout programs aim to create a comprehensive resource of targeted mutations for all protein-coding genes.
Existing methods require optimization for high-throughput generation of diverse alleles.

Purpose of the Study:

To establish a high-throughput gene-targeting pipeline for generating reporter-tagged, conditional alleles.
To scale up the production of targeted mutations for a wider research community.
To lay the foundation for genome-wide efforts to decipher mammalian gene function.

Main Methods:

Development of a high-throughput pipeline integrating computational allele design.
Utilizing 96-well modular vector construction for efficient allele generation.
Employing high-efficiency gene-targeting strategies in germline-competent C57BL/6N embryonic stem cells.

Main Results:

Successful establishment of a scalable gene-targeting pipeline.
Generation of over 12,000 vectors and 9,000 conditional targeted alleles.
Demonstration of high-throughput genome engineering capabilities.

Conclusions:

The developed pipeline enables unprecedented scale in generating conditional targeted alleles.
This high-throughput genome engineering approach is applicable to rat and human stem cells.
The resource facilitates future genome-wide studies to determine the function of all mammalian genes.