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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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Related Experiment Video

Updated: Jul 27, 2025

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Published on: May 15, 2018

Genome-scale targeted mutagenesis in

Jianjie He^1,2, Kai Zhang^1,2, Shuxiang Yan^1,2

¹Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.

Genome Research

|June 8, 2023

Summary

This summary is machine-generated.

This study introduces a pooled CRISPR library for efficient genome-scale targeted editing in the allotetraploid crop Brassica napus. This method advances functional genomics and crop breeding in polyploid plants.

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Last Updated: Jul 27, 2025

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

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Published on: May 15, 2018

Optogenetic Random Mutagenesis Using Histone-miniSOG in C. elegans

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Published on: November 14, 2016

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Published on: January 26, 2017

Area of Science:

Plant genomics
Biotechnology
Crop science

Background:

CRISPR-Cas9 technology has enabled large-scale functional genomics in diploid crops.
Targeted mutagenesis in complex polyploid genomes remains a significant challenge.

Purpose of the Study:

To demonstrate the feasibility of a pooled CRISPR library for genome-scale targeted editing in the allotetraploid Brassica napus.
To establish a high-throughput method for reverse genetic screening in polyploid plants.

Main Methods:

Designed 18,414 sgRNAs targeting 10,480 genes in Brassica napus.
Generated 1104 transgenic plants and analyzed gene editing efficiency.
Investigated Cas9 cleavage patterns at target sites.

Main Results:

Achieved a 52.2% mutation rate in 178 analyzed genes.
Discovered that Cas9 cleaves all target sites guided by a single sgRNA in polyploids.
Identified novel genes influencing fatty acid profile and seed oil content.

Conclusions:

Pooled CRISPR libraries are effective for high-throughput targeted mutagenesis in allotetraploid crops.
This approach provides valuable resources for Brassica napus functional genomics and breeding.
Offers a reference for polyploid plant research and crop improvement.