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Published online 28 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2336-2341
© 2004 Oxford University Press

A novel replicating circular DNAzyme

Fei Chen, Ruijian Wang, Zhe Li, Bin Liu, Xiaoping Wang, Yanhong Sun, Dongyun Hao and Jin Zhang*

Key Lab for Molecular Enzymology and Engineering of the Ministry of Education (Jilin University), Changchun, 130023, P.R. China

*To whom correspondence should be addressed. Tel/Fax: +86 431 8980440; Email: zhangjin{at}jlu.edu.cn
Correspondence may also be addressed to Dongyun Hao. Tel/Fax: +86 431 8980440; Email: hao{at}jlu.edu.cn

Received January 10, 2004; Revised and Accepted March 25, 2004

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 ß-lactamase mRNA. These circular DNAzymes showed in vitro catalytic efficiencies (kcat/KM) of 7.82 x 106 and 1.36 x 107 M–1·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 ß-lactamase and bacterial growth. This study thus provides a novel strategy to produce replicating DNAzymes which may find widespread applications.


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