A novel replicating circular DNAzyme.
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.
Chen
Fei
F
Key Lab for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun, 130023, P.R. China.
Wang
Ruijian
R
Li
Zhe
Z
Liu
Bin
B
Wang
Xiaoping
X
Sun
Yanhong
Y
Hao
Dongyun
D
Zhang
Jin
J
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
D002413:Cations, Divalent; D021881:DNA, Catalytic; D004270:DNA, Circular; D001618:beta-Lactamases; D008274:Magnesium