title: A novel replicating circular DNAzyme.
abstract: 10-23 DNAzyme has the potential to suppress gene expressions through sequence-specific mRNA cleavage. However, the dependence on exogenous delivery limits its applications. The objective of this work is to establish a replicating DNAzyme in bacteria using a single-stranded DNA vector. By cloning the 10-23 DNAzyme into the M13mp18 vector, we constructed two circular DNAzymes, C-Dz7 and C-Dz482, targeting the beta-lactamase mRNA. These circular DNAzymes showed in vitro catalytic efficiencies (kcat/K(M)) of 7.82 x 10(6) and 1.36 x 10(7) M(-1) x min(-1), respectively. Their dependence on divalent metal ions is similar to that found with linear 10-23 DNAzyme. Importantly, the circular DNAzymes were not only capable of replicating in bacteria but also exhibited high activities in inhibiting beta-lactamase and bacterial growth. This study thus provides a novel strategy to produce replicating DNAzymes which may find widespread applications.
authors.affiliation: Key Lab for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, 130023, P.R. China.
authors.forename: Fei
authors.initials: F
authors.lastname: Chen
authors.forename: Ruijian
authors.initials: R
authors.lastname: Wang
authors.forename: Zhe
authors.initials: Z
authors.lastname: Li
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Wang
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Sun
authors.forename: Dongyun
authors.initials: D
authors.lastname: Hao
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D002384:Catalysis / Q000187:drug effects; D002413:Cations, Divalent / Q000494:pharmacology; D021881:DNA, Catalytic / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D004270:DNA, Circular / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics; D008274:Magnesium / Q000494:pharmacology; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D021881:DNA, Catalytic; D004270:DNA, Circular; D001618:beta-Lactamases; D008274:Magnesium