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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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Simultaneous detection of multiple transgenes for genetically-modified mouse strains.

Hatsumi Nakata¹, Tomomi Hashimoto, Yukiko Seki

¹Experimental Animal Division, RIKEN BioResource Center, Koyadai,Tsukuba, Ibaraki, Japan.

Experimental Animals

|August 6, 2009

Summary

RIKEN BRC developed KO-survey, a simultaneous PCR test for efficiently verifying genetic modifications in mouse strains. This method ensures high-quality genetically-modified mice and accurate documentation for research purposes.

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Generation of Genetically Modified Mice through the Microinjection of Oocytes

Generation of Genetically Modified Mice through the Microinjection of Oocytes

Published on: June 15, 2017

Area of Science:

Genetics
Animal Models
Molecular Biology

Background:

Maintaining high-quality genetically-modified mice (transgenic, knockout, conditional knockout) is challenging due to scientific and legal requirements.
RIKEN BRC houses a diverse collection of genetically-modified mouse strains.
Accurate and rapid verification of genetic modifications is crucial for research integrity.

Purpose of the Study:

To establish an efficient method for verifying genetic modifications in mouse strains.
To ensure the genetic quality and accurate characterization of mouse models.
To support researchers with reliable genetically-modified mouse lines.

Main Methods:

Development of a simultaneous Polymerase Chain Reaction (PCR) test named "KO-survey".
Optimization of PCR conditions to detect multiple transgene markers.
Division of detection into two sets: Set I (neo, puro, pgk-neo, lacZ, HSVtk-neo) and Set II (hyg, IRES, cre, flp, Gfp).

Main Results:

The "KO-survey" enables simultaneous detection of various transgenes.
Optimized PCR conditions allow for quick and thorough examination of genetic modifications.
The assay effectively distinguishes between different genetic modifications in mouse strains.

Conclusions:

The "KO-survey" PCR test is a valuable tool for quality control of genetically-modified mouse strains.
This method enhances the reliability of genetic information provided with mouse models.
RIKEN BRC can ensure the provision of high-quality, accurately characterized mouse strains to users.