title: An alternative novel tool for DNA editing without target sequence limitation: the structure-guided nuclease.
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.
authors.affiliation: Department of Pharmacology, Jinling Hospital, School of Medicine, Nanjing University, No. 305 Zhongshan East Road, Nanjing, 210002, People's Republic of China.
authors.forename: Shu
authors.initials: S
authors.lastname: Xu
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Shasha
authors.initials: S
authors.lastname: Cao
authors.affiliation: Department of Pharmacology, Jinling Hospital, School of Medicine, Nanjing University, No. 305 Zhongshan East Road, Nanjing, 210002, People's Republic of China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Zou
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Yunyun
authors.initials: Y
authors.lastname: Yue
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Chun
authors.initials: C
authors.lastname: Gu
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Pei
authors.initials: P
authors.lastname: Wang
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Dong
authors.affiliation: Department of Pharmacology, Jinling Hospital, School of Medicine, Nanjing University, No. 305 Zhongshan East Road, Nanjing, 210002, People's Republic of China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Xiang
authors.affiliation: College of Pharmaceutical Science, Soochow University, No. 199, Renai Road, Suzhou, 215123, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Li
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China. zhums@nicemice.cn.
authors.forename: Minsheng
authors.initials: M
authors.lastname: Zhu
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Nanjing University, 12 Xuefu Road, Pukou High-tech Development Zone, Nanjing, 210061, People's Republic of China. qingshun@nju.edu.cn.
authors.forename: Qingshun
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, Jinling Hospital, School of Medicine, Nanjing University, No. 305 Zhongshan East Road, Nanjing, 210002, People's Republic of China. ghzhou@nju.edu.cn.
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045585:Flap Endonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000072669:Gene Editing; D018390:Gene Targeting / Q000379:methods; D008954:Models, Biological; D009154:Mutation; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D015027:Zebrafish
chemical_list: D004277:DNA, Single-Stranded; D004247:DNA; D004706:Endodeoxyribonucleases; D045585:Flap Endonucleases; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific
keywords: DNA editing; Fok I; Guide DNA; Structure-guided endonuclease; flap endonuclease-1 (FEN-1)