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Precise A•T to G•C base editing in the zebrafish genome
Wei Qin1, Xiaochan Lu2, Yunxing Liu2
1State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, 518055, China. qinwei@pkusz.edu.cn.
BMC Biology
|November 22, 2018
Summary
We optimized adenine base editors (ABEs) for zebrafish, creating a new tool (zABE7.10) for efficient A•T to G•C gene editing. This system successfully models human diseases caused by point mutations.
Area of Science:
- Genetics
- Molecular Biology
- Zebrafish Models
Background:
- Base editors enable precise DNA modifications without double-strand breaks.
- Adenine base editors (ABEs) convert A•T to G•C in various organisms.
- Investigating ABE efficiency in zebrafish is crucial for disease modeling.
Purpose of the Study:
- To assess and optimize adenine base editor activity in zebrafish.
- To establish a robust system for A•T to G•C base editing in zebrafish.
- To develop new zebrafish models for human genetic diseases.
Main Methods:
- Codon optimization of ABE7.10 for zebrafish.
- Testing zABE7.10 activity across multiple genomic loci.
- Introducing a specific point mutation in rps14 to model 5q-Syndrome.
- Enhancing editing efficiency using bipartite nuclear localization signals (bpNLS).
Main Results:
- Codon-optimized zABE7.10 showed high activity and germline targeting efficiency in zebrafish.
- A zebrafish model for 5q-Syndrome was successfully created.
- bpNLS modification improved ABE-mediated editing efficiency by 1.96-fold on average.
Conclusions:
- The zABE7.10 system enables efficient A•T to G•C base editing in zebrafish.
- This tool enhances the capacity for modeling human diseases in zebrafish.
- Optimized base editing provides a powerful strategy for genetic research in zebrafish.
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