PUBLICATION
Precise A•T to G•C base editing in the zebrafish genome
- Authors
- Qin, W., Lu, X., Liu, Y., Bai, H., Li, S., Lin, S.
- ID
- ZDB-PUB-181127-28
- Date
- 2018
- Source
- BMC Biology 16: 139 (Other)
- Registered Authors
- Lin, Shuo
- Keywords
- Adenine base editors, Base editing, Zebrafish
- MeSH Terms
-
- Animals
- Base Sequence
- Fish Proteins
- Gene Editing/methods*
- Genome*
- Point Mutation*
- Zebrafish/genetics*
- PubMed
- 30458760 Full text @ BMC Biol.
Citation
Qin, W., Lu, X., Liu, Y., Bai, H., Li, S., Lin, S. (2018) Precise A•T to G•C base editing in the zebrafish genome. BMC Biology. 16:139.
Abstract
Background Base editors are a class of genome editing tools with the ability to efficiently induce point mutations in genomic DNA, without inducing double-strand breaks or relying on homology-direct repair as in other such technologies. Recently, adenine base editors (ABEs) have been developed to mediate the conversion of A•T to G•C in genomic DNA of human cells, mice, and plants. Here, we investigated the activity and efficiency of several adenine base editors in zebrafish and showed that base editing can be used to create new models of pathogenic diseases caused by point mutations.
Results The original ABE7.10 exhibits almost no activity in zebrafish. After codon optimization, we found that a zABE7.10 variant could induce targeted conversion of adenine to guanine in zebrafish at multiple tested genomic loci, and all the target sites showed a high rate of germline targeting efficiency. Furthermore, using this system, we established a zebrafish model of 5q-Syndrome that contained a new point mutation in rps14. The further modification of zABE7.10 by a bipartite nuclear localization signals (bpNLS) resulted in 1.96-fold average improvement in ABE-mediated editing efficiency at four sites.
Conclusions Collectively, this system, designated as zABE7.10, provides a strategy to perform A•T to G•C base editing in zebrafish and enhances its capacity to model human diseases.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping