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
  • Animals
  • Base Sequence
  • DNA/chemistry
  • DNA/genetics*
  • DNA/metabolism
  • DNA, Single-Stranded/genetics
  • DNA, Single-Stranded/metabolism
  • Deoxyribonucleases, Type II Site-Specific/chemistry
  • Deoxyribonucleases, Type II Site-Specific/genetics
  • Deoxyribonucleases, Type II Site-Specific/metabolism
  • Endodeoxyribonucleases/chemistry
  • Endodeoxyribonucleases/genetics
  • Endodeoxyribonucleases/metabolism*
  • Flap Endonucleases/chemistry
  • Flap Endonucleases/genetics
  • Flap Endonucleases/metabolism
  • Gene Editing*
  • Gene Targeting/methods
  • Models, Biological
  • Mutation
  • Sequence Analysis, DNA
  • Substrate Specificity
  • Zebrafish
PubMed
27634179 Full text @ Genome Biol.
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
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping