A novel replicating circular DNAzyme
A novel replicating circular DNAzyme
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DOI：
10.1093/nar/gkh547
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发表时间：
2004-04-01
影响因子：
14.9
通讯作者：
Zhang, J
中科院分区：
文献类型：
--
作者：
Chen, F;Wang, RJ;Zhang, J
10-23 DNAzyme具有通过序列特异性mRNA切割来抑制基因表达的潜力。然而，对外源性递送的依赖限制了其应用。本工作的目的是使用单链DNA载体在细菌中建立可复制的DNAzyme。通过将10-23 DNA酶克隆到M13 mp 18载体中，我们构建了两种针对β-内酰胺酶mRNA的环状DNA酶C-Dz（7）和C-Dz（482）。这些环状DNA酶的体外催化效率（k（cat）/K-M）分别为7.82 × 10（6）和1.36 × 10（7）M-1.min-1。它们对二价金属离子的依赖性与线性10-23 DNAzyme相似。重要的是，环状DNA酶不仅能够在细菌中复制，而且在抑制β-内酰胺酶和细菌生长方面表现出高活性。因此，这项研究提供了一种新的策略，生产复制的DNA酶，可能会发现广泛的应用。
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-Dz(7) and C-Dz(482), targeting the beta-lactamase mRNA. These circular DNAzymes showed in vitro catalytic efficiencies (k(cat)/K-M) of 7.82 x 10(6) and 1.36 x 10(7) 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 beta-lactamase and bacterial growth. This study thus provides a novel strategy to produce replicating DNAzymes which may find widespread applications.