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Genome Editing of Mouse by Cytoplasmic Injection
1Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi, 371-8512, Gunma, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|June 24, 2017
Summary
This study details a method for creating genome-edited knockout mice using CRISPR/Cas9 technology. The guide focuses on cytoplasmic injection of Cas9 and guide RNA into fertilized eggs for efficient gene modification.
Area of Science:
- Molecular Biology
- Genetics
- Animal Models
Background:
- CRISPR/Cas technology offers a powerful tool for precise genome editing.
- Generating genetically modified animal models is crucial for biological research.
- Various methods exist for introducing CRISPR/Cas components into animal zygotes.
Purpose of the Study:
- To provide a detailed instructional guide for generating knockout mice.
- To demonstrate the efficacy of cytoplasmic injection for CRISPR/Cas delivery.
- To facilitate the rapid production of genome-edited animal models.
Main Methods:
- Utilizing CRISPR/Cas9 system for genome editing.
- Employing in vitro transcribed Cas9 and guide RNA (gRNA).
- Performing cytoplasmic injection of Cas9/gRNA into fertilized mouse eggs.
Main Results:
- Successful generation of knockout mice using the described method.
- Demonstration of efficient gene editing via cytoplasmic injection.
- Establishment of a reproducible protocol for genome editing in mice.
Conclusions:
- Cytoplasmic injection is an effective method for delivering CRISPR/Cas9 components.
- This protocol enables the rapid generation of genome-edited knockout mice.
- The developed method serves as a valuable resource for researchers in genetics and molecular biology.
