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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.

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Updated: Sep 24, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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Published on: December 11, 2020

High-Throughput Gene Mutagenesis Screening Using Base Editing.

Philippe C Després^1,2,3,4, Alexandre K Dubé^5,6,7,8,9, Nozomu Yachie^10,11,12,13

¹Département de Biochimie, Microbiologie et Bio-informatique, Faculté de Sciences et Génie, Université Laval, Québec, QC, Canada. philippe.despres.3@ulaval.ca.

Methods in Molecular Biology (Clifton, N.J.)

|May 6, 2022

Summary

Large-scale base editing screens in yeast enable thousands of simultaneous mutations using the Target-AID editor. This method efficiently measures mutation effects on fitness without double-stranded DNA breaks.

Keywords:

Base editing CRISPR-Cas9 Functional genomics High-throughput Mutagenesis Systems biology Target-AID Yeast

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Published on: April 8, 2017

Area of Science:

Genetics and Genomics
Molecular Biology
Biotechnology

Background:

CRISPR-Cas9 technology offers precise genome engineering.
Base editing allows targeted nucleotide changes without double-stranded breaks.
Efficient screening methods are crucial for understanding gene function.

Purpose of the Study:

To develop and implement large-scale base editing screens in yeast.
To utilize the Target-AID base editor for simultaneous mutagenesis.
To assess the impact of numerous mutations on cellular fitness.

Main Methods:

Employed the Target-AID base editor for programmable mutagenesis in yeast.
Designed and executed pooled growth competition assays.
Utilized DNA sequencing of guide RNAs (gRNAs) as barcodes to track mutations.

Main Results:

Successfully performed large-scale base editing screens in yeast.
Demonstrated the ability to mutate thousands of genomic sites simultaneously.
Quantified the fitness effects of these mutations through competition assays.

Conclusions:

Large-scale base editing screens are feasible and effective in yeast.
This approach provides a powerful tool for functional genomics and understanding mutation impacts.
The Target-AID editor facilitates efficient, multiplexed genome engineering for fitness studies.