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Updated: Oct 10, 2025
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Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
Published on: December 16, 2022
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The versatile electric condition in mouse embryos for genome editing using a three-step square-wave pulse
Kenta Nakano1, Yukiko Shimizu1, Tetsuya Arai1
1Department of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine (NCGM), 1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan.
Experimental Animals
|December 9, 2021
Summary
The Technique for Animal Knockout system by Electroporation (TAKE) offers a simple, efficient method for generating genetically modified mice using CRISPR/Cas9. Optimized electric conditions enable rapid gene editing in various mouse strains, facilitating research on human diseases and gene functions.
Area of Science:
- * Molecular Biology and Genetics
- * Animal Models and Gene Editing
Background:
- * Generating genetically modified (GM) mice is crucial for studying gene function and human diseases.
- * CRISPR/Cas9 technology has revolutionized gene editing but requires efficient delivery methods.
- * Electroporation offers a potentially simpler alternative to microinjection for delivering gene-editing components.
Purpose of the Study:
- * To optimize electroporation electric conditions for efficient gene editing in mouse embryos.
- * To validate the versatility of the optimized method across different inbred mouse strains.
- * To establish a rapid and efficient protocol for generating genome-edited mice.
Main Methods:
- * Optimization of electroporation pulse settings (pore-forming and transfer pulses) for vitrified-warmed mouse embryos.
- * Application of optimized conditions to fresh embryos from C57BL/6NCr, BALB/cCrSlc, FVB/NJcl, and C3H/HeJJcl strains.
- * Utilizing a 5 mm electrode gap for high-throughput embryo processing (over 100 embryos/5 min).
Main Results:
- * Identified optimal electric pulse settings for efficient delivery of gRNA/Cas9 complex into zygotes.
- * Successfully generated GM mice across multiple widely used inbred strains without zona pellucida thinning.
- * Achieved high embryo developmental rates and genome-editing efficiency in offspring.
Conclusions:
- * The optimized electroporation technique (TAKE) provides a versatile, rapid, and efficient method for generating GM mice.
- * This approach simplifies the generation of genome-edited mice, reducing technical barriers.
- * Facilitates research into human diseases and gene functions through easier access to custom mouse models.
