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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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Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

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Targeted Mutagenesis Using RNA-guided Endonucleases in Mosses.

Toshihisa Nomura¹, Hitoshi Sakakibara^1,2

¹RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro, Tsurumi, Yokohama, Japan.

|September 20, 2021

Summary

This study presents a straightforward RNA-guided endonuclease (RGEN) method for targeted genome editing in the model moss Physcomitrella patens. This technique enables efficient mutagenesis and genotyping, applicable to diverse moss species.

Keywords:

CRISPR/Cas system Genome editing Mosses RNA-guided endonucleases Targeted mutagenesis

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Last Updated: Oct 19, 2025

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Published on: February 5, 2019

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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Published on: March 25, 2020

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

Area of Science:

Plant Biology
Genetics
Molecular Biology

Background:

Genome editing technologies are crucial for understanding plant gene function.
RNA-guided endonucleases (RGENs) offer precise DNA modification capabilities.
Efficient genome editing tools are needed for diverse plant species, including non-model organisms.

Purpose of the Study:

To develop a simple and effective method for targeted mutagenesis and genotyping in mosses using RGENs.
To demonstrate the applicability of the RGEN system in both a model moss (Physcomitrella patens) and a non-model moss (Scopelophilla cataractae).

Main Methods:

Utilized RNA-guided endonucleases (RGENs) for targeted DNA modification.
Applied the RGEN system for mutagenesis and genotyping in Physcomitrella patens.
Adapted the RGEN method for targeted mutagenesis in Scopelophilla cataractae.

Main Results:

Successfully demonstrated targeted mutagenesis and genotyping in the model moss Physcomitrella patens.
Achieved targeted mutagenesis in the non-model moss Scopelophilla cataractae using a similar RGEN approach.
Validated the broad applicability of the developed RGEN system across different moss species.

Conclusions:

A simple RGEN-based method facilitates genome editing in mosses.
This system is effective for both model and non-model moss species.
The RGEN approach holds potential for wide application in bryophyte research.