PUBLICATION
An alternative novel tool for DNA editing without target sequence limitation: the structure-guided nuclease
- Authors
- Xu, S., Cao, S., Zou, B., Yue, Y., Gu, C., Chen, X., Wang, P., Dong, X., Xiang, Z., Li, K., Zhu, M., Zhao, Q., Zhou, G.
- ID
- ZDB-PUB-160917-7
- Date
- 2016
- Source
- Genome biology 17: 186 (Journal)
- Registered Authors
- Zhao, Qingshun
- Keywords
- DNA editing, Fok I, Guide DNA, Structure-guided endonuclease, flap endonuclease-1 (FEN-1)
- MeSH Terms
-
- DNA/chemistry
- DNA/genetics*
- DNA/metabolism
- Models, Biological
- Substrate Specificity
- Sequence Analysis, DNA
- Flap Endonucleases/chemistry
- Flap Endonucleases/genetics
- Flap Endonucleases/metabolism
- Endodeoxyribonucleases/chemistry
- Endodeoxyribonucleases/genetics
- Endodeoxyribonucleases/metabolism*
- Deoxyribonucleases, Type II Site-Specific/chemistry
- Deoxyribonucleases, Type II Site-Specific/genetics
- Deoxyribonucleases, Type II Site-Specific/metabolism
- DNA, Single-Stranded/genetics
- DNA, Single-Stranded/metabolism
- Base Sequence
- Mutation
- Animals
- Zebrafish
- Gene Targeting/methods
- Gene Editing*
- PubMed
- 27634179 Full text @ Genome Biol.
Citation
Xu, S., Cao, S., Zou, B., Yue, Y., Gu, C., Chen, X., Wang, P., Dong, X., Xiang, Z., Li, K., Zhu, M., Zhao, Q., Zhou, G. (2016) An alternative novel tool for DNA editing without target sequence limitation: the structure-guided nuclease. Genome biology. 17:186.
Abstract
Engineered endonucleases are a powerful tool for editing DNA. However, sequence preferences may limit their application. We engineer a structure-guided endonuclease (SGN) composed of flap endonuclease-1 (FEN-1), which recognizes the 3' flap structure, and the cleavage domain of Fok I (Fn1), which cleaves DNA strands. The SGN recognizes the target DNA on the basis of the 3' flap structure formed between the target and the guide DNA (gDNA) and cut the target through its Fn1 dimerization. Our results show that the SGN, guided by a pair of gDNAs, cleaves transgenic reporter gene and endogenous genes in zebrafish embryonic genome.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping