Genetic encoding of DNA nanostructures and their self-assembly in living bacteria.
Genetic encoding of DNA nanostructures and their self-assembly in living bacteria.
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DOI：
10.1038/ncomms11179
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发表时间：
2016-04-19
影响因子：
16.6
通讯作者：
Voigt CA
中科院分区：
文献类型：
--
作者：
Elbaz J;Yin P;Voigt CA
DNA纳米技术领域已经利用DNA碱基配对的可编程性来指导单链DNA（ssDNA）组装成所需的3D结构。在这里，我们展示了在大肠杆菌中表达ssDNA（32-205 nt）的能力，其可以在体内形成结构或被纯化用于体外组装。每个ssDNA由一个基因编码，该基因转录成含有3′-发夹（HTBS）的非编码RNA。HTBS招募HIV逆转录酶，其使DNA合成成核并通过鼠白血病逆转录酶辅助延伸。体内产生的纯化的ssDNA用于在体外组装大的1D线（300 nm）和2D片（5.8 μm2）。细胞内组装证明使用四ssDNA交叉纳米结构，招聘分裂YFP时，正确组装。遗传编码DNA纳米结构为它们的生产以及在活细胞中的应用提供了途径。 DNA纳米结构有潜力成为生物学许多领域的强大工具，但它们很难完全在体内制造。在这里，作者将联合收割机RNA发夹和逆转录结合起来，在细胞环境中产生并组装一个复杂的DNA结构。
The field of DNA nanotechnology has harnessed the programmability of DNA base pairing to direct single-stranded DNAs (ssDNAs) to assemble into desired 3D structures. Here, we show the ability to express ssDNAs in Escherichia coli (32–205 nt), which can form structures in vivo or be purified for in vitro assembly. Each ssDNA is encoded by a gene that is transcribed into non-coding RNA containing a 3′-hairpin (HTBS). HTBS recruits HIV reverse transcriptase, which nucleates DNA synthesis and is aided in elongation by murine leukemia reverse transcriptase. Purified ssDNA that is produced in vivo is used to assemble large 1D wires (300 nm) and 2D sheets (5.8 μm2) in vitro. Intracellular assembly is demonstrated using a four-ssDNA crossover nanostructure that recruits split YFP when properly assembled. Genetically encoding DNA nanostructures provides a route for their production as well as applications in living cells. DNA nanostructures have the potential to be powerful tools in many areas of biology however they are difficult to manufacture completely in vivo. Here the authors combine RNA hairpins and reverse transcription to generate and assemble a complex DNA structure inside the cellular environment.