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Rapid and Efficient Spatiotemporal Monitoring of Normal and Aberrant Cytosine Methylation within Intact Zebrafish Embryos
Published on: August 18, 2022
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In vivo DNA methylation editing in zebrafish
Fang Liang1, Zijiong Dong1, Jianmin Ye1
1Institute of Modern Aquaculture Science and Engineering, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Sciences, South China Normal University, Guangzhou, P. R. China.
Epigenetics
|March 22, 2023
Summary
This study introduces CRISPR/dCas9 epigenome editing for zebrafish (Danio rerio), enabling targeted DNA methylation changes. This novel technique provides a powerful new tool for genetic research in this important biomedical model organism.
Area of Science:
- Epigenetics
- Molecular Biology
- Zebrafish Model Organisms
Background:
- CRISPR/dCas9 epigenome editing is a powerful technique for targeted DNA modification.
- CRISPR/dCas9 fused with catalytic domains (DnmtCD or TetCD) enables DNA methylation editing.
- Zebrafish (Danio rerio) are crucial biomedical models, but lack established CRISPR-based DNA methylation editing tools.
Purpose of the Study:
- To demonstrate the feasibility of CRISPR/dCas9-based DNA methylation editing in zebrafish.
- To establish CRISPR/dCas9-Dnmt7 and CRISPR/dCas9-Tet2 catalytic domain fusions for targeted epigenetic modification in vivo.
- To evaluate the potential of these systems as a toolkit for DNA methylation editing in zebrafish.
Main Methods:
- Utilized CRISPR/dCas9 fused with Dnmt7 and Tet2 catalytic domains.
- Applied these fusions for site-specific genomic DNA methylation editing in vivo in zebrafish.
- Validated the targeted editing of DNA methylation patterns.
Main Results:
- Successfully demonstrated site-specific genomic DNA methylation editing in vivo in zebrafish.
- Established CRISPR/dCas9-Dnmt7 and CRISPR/dCas9-Tet2 catalytic domain fusions as functional tools in zebrafish.
- Showcased the ability to manipulate DNA methylation at targeted genomic loci.
Conclusions:
- CRISPR/dCas9-Dnmt7 and -Tet2 catalytic domain fusions are effective for in vivo DNA methylation editing in zebrafish.
- This represents the first report of CRISPR-based epigenome editing in zebrafish.
- These systems offer a promising and efficient toolkit for DNA methylation research and disease modeling in zebrafish.
