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Construction and characterization of metal ion-containing DNA nanowires for synthetic biology and nanotechnology

机译:用于合成生物学和纳米技术的含金属离子的DNA纳米线的构建和表征

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DNA is an attractive candidate for integration into nanoelectronics as a biological nanowire due to its linear geometry, definable base sequence, easy, inexpensive and non-toxic replication and self-assembling properties. Recently we discovered that by intercalating Ag+ in polycytosine-mismatch oligonucleotides, the resulting C-Ag+-C duplexes are able to conduct charge efficiently. To map the functionality and biostability of this system, we built and characterized internally-functionalized DNA nanowires through non-canonical, Ag+-mediated base pairing in duplexes containing cytosine-cytosine mismatches. We assessed the thermal and chemical stability of ion-coordinated duplexes in aqueous solutions and conclude that the C-Ag+-C bond forms DNA duplexes with replicable geometry, predictable thermodynamics, and tunable length. We demonstrated continuous ion chain formation in oligonucleotides of 11–50 nucleotides (nt), and enzyme ligation of mixed strands up to six times that length. This construction is feasible without detectable silver nanocluster contaminants. Functional gene parts for the synthesis of DNA- and RNA-based, C-Ag+-C duplexes in a cell-free system have been constructed in an Escherichia coli expression plasmid and added to the open-source BioBrick Registry, paving the way to realizing the promise of inexpensive industrial production. With appropriate design constraints, this conductive variant of DNA demonstrates promise for use in synthetic biological constructs as a dynamic nucleic acid component and contributes molecular electronic functionality to DNA that is not already found in nature. We propose a viable route to fabricating stable DNA nanowires in cell-free and synthetic biological systems for the production of self-assembling nanoelectronic architectures.
机译:由于其线性的几何形状,可定义的碱基序列,简单,廉价且无毒的复制和自组装特性,DNA是作为生物纳米线整合到纳米电子学中的诱人候选物。最近我们发现,通过在多胞嘧啶错配寡核苷酸中插入Ag + ,所得的C-Ag + -C双链体能够有效地传导电荷。为了绘制该系统的功能和生物稳定性,我们通过非规范的,Ag + 介导的碱基对在含有胞嘧啶-胞嘧啶错配的双链体中构建并表征了内部功能化的DNA纳米线。我们评估了离子配位双链体在水溶液中的热稳定性和化学稳定性,并得出结论,C-Ag + -C键形成具有可复制几何形状,可预测的热力学和可调长度的DNA双链体。我们证明了在11–50个核苷酸(nt)的寡核苷酸中连续的离子链形成,混合链的酶连接长达该长度的六倍。没有可检测的银纳米团簇污染物,这种构造是可行的。已在大肠杆菌表达质粒中构建了用于在无细胞系统中合成基于DNA和RNA的C-Ag + -C双链体的功能基因部分,并将其添加到开源中BioBrick Registry,为实现廉价工业生产的承诺铺平了道路。在适当的设计约束下,DNA的这种导电变异体显示出有望在合成生物构建物中用作动态核酸成分,并为自然界中尚未发现的DNA贡献分子电子功能。我们提出了一条可行的途径,可以在无细胞和合成生物系统中制造稳定的DNA纳米线,以生产自组装的纳米电子体系结构。

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