Engineering a DNAzyme-Based Operon System for the Production of DNA Nanoscaffolds in Living Bacteria
Engineering a DNAzyme-Based Operon System for the Production of DNA Nanoscaffolds in Living Bacteria
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DOI：
10.1021/acssynbio.9b00415
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发表时间：
2020-02-01
影响因子：
4.7
通讯作者：
Elbaz, Johann
中科院分区：
文献类型：
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作者：
Alon, Dan M.;Voigt, Christopher A.;Elbaz, Johann
使用DNA在活细胞中创建纳米支架的能力有可能成为合成生物学中的强大工具。然而，到目前为止，遗传编码的DNA纳米结构仅限于简单的结构，因为缺乏可以在单个细菌中产生多个单链DNA的遗传部分。在这里，我们开发了一个系统，通过使用一个模仿操纵子的单一寡核苷酸基因来克服这一挑战。这是通过将非编码的RNA转化为使用R3-DNAzyme(基于DNAzyme的操纵子)自切割成多个单链DNA来实现的。我们展示了应用基于DNAzyme的操纵子来产生四个单链DNA交叉纳米结构(25 Nm)的能力，当适当组装时，该结构可以招募分裂的YFP。该系统能够在体内形成更复杂的DNA纳米结构，从而为进一步将DNA纳米技术领域整合到基础生物学、生物工程和医学应用的活细菌中铺平了道路。
The ability to create nanoscaffolds within living cells using DNA has the potential to become a powerful tool in synthetic biology. However, to date, genetically encoded DNA nanostructures are limited to simple architecture due to the lack of genetic parts that can produce multiple ssDNAs in a single bacterium. Here, we develop a system that overcomes this challenge by using a single oligo gene mimicking operons. This was achieved by converting a noncoding RNA into a long ssDNA that self-cleaves into multiple ssDNAs using R3-DNAzymes (DNAzyme-based operon). We demonstrate the ability to apply the DNAzyme-based operon to produce a four-ssDNA crossover nanostructure (25 nm) that recruits split YFPs when properly assembled. This system enables the formation of more complex DNA nanostructures in vivo and thus paves the way to further integrate the field of DNA nanotechnology into living bacteria for basic biology, bioengineering, and medicine applications.