Thio-Ether Nucleic Acids: Clicking Together Synthetic Poly(Nucleic Acids)

描述（由申请人提供）：DNA代表最有能力和最强大的生物分子结构之一：作为遗传物质起作用，与互补链杂交，作为适体起作用，在转录和翻译中具有活性，以及能够诱导组织并能够形成设计的纳米结构。DNA执行许多天然生物学功能以及用于药物开发、生物检测、基因敲除和基因组学测定以及许多其他。尽管有这些能力，DNA仍然非常昂贵和/或难以实现用于除了最有价值的应用之外的任何应用，特别是在材料领域。所提出的研究的总体假设是，基于硫醇-烯的点击化学与寡核苷酸合成的协同组合将导致形成全新的和高能力的一类寡核苷酸结构，即，硫醚核酸（TNA），其能够在体内、体外和生物功能应用如生物检测中以序列特异性与DNA相互作用，并作为适体模拟物。硫醇-烯点击化学与合成寡核苷酸生产的协同组合将产生具有增强的DNA结合能力的功能性寡核苷酸，更容易大规模生产，并且具有优于DNA和PNA结构的独特化学优势。概括地说，该项目的总体目标是开发和执行一个全新的类别的寡核苷酸分子的初步表征，这将具有显着的优势，比生物寡核苷酸（DNA和RNA）以及合成的PNA的DNA杂交，生物检测，和其他生物功能的应用。这项工作的具体目标是（i）合成碱基功能化的TNA单体，（ii）从这些单体形成受控序列寡聚体，并评估它们与DNA的强选择性杂交的能力，以及（iii）证明这些TNA序列在KRAS癌基因SNP检测中的实用性。

DESCRIPTION (provided by applicant): DNA represents one of the most capable and powerful biomolecular structures: functional as genetic material, hybridizing to complementary strands, being functional as aptamers, and active in transcription and translation as well as being capable of inducing organization and enabling the formation of designed nanostructures. DNA performs numerous natural biological functions as well as being used in drug development, biodetection, gene knockout, and genomics assays as well as many others. Despite these capabilities, DNA remains prohibitively expensive and/or difficult to implement for use in any but the most valuable applications, particularly in the materials realm. The overall hypothesis of the proposed research is that the synergistic combination of thiol-ene-based click chemistry with oligonucleotide synthesis will lead to the formation of an entirely new and highly capable class of oligonucleotide structures, i.e., thio-ether nucleic acids (TNAs), that are capable of interacting with sequence specificity with DNA in vivo, in vitro and in biofunctional applications such as biodetection and as aptamer mimics. The synergistic combination of thiol-ene click chemistry with synthetic oligonucleotide production will yield functional oligonucleotides that have enhanced DNA-binding capability, are easier to produce at scale, and have distinct chemical advantages over DNA and PNA structures. Broadly, the overall aim of this project is to develop and perform preliminary characterization of an entirely new class of oligonucleotide molecules that would have significant advantages over both biological oligonucleotides (DNA and RNA) as well as the synthetic PNAs for DNA hybridization, for biodetection, and for other biofunctional applications. The specific aims of this work are to (i) synthesize base-functionalized TNA monomers, (ii) form controlled sequence oligomers from these monomers and evaluate their capacity for strong, selective hybridization with DNA, and (iii) demonstrate the utility of these TNA sequences in SNP detection of the KRAS oncogene.