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

描述(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：作为遗传物质的功能，与互补的链杂交，作为适配子的功能，活跃于转录和翻译，以及能够诱导组织和使设计的纳米结构的形成。DNA具有许多天然的生物学功能，并被用于药物开发、生物检测、基因敲除和基因组分析等许多领域。尽管有这些能力，DNA仍然昂贵和/或难以实现，用于除最有价值的应用之外的任何应用，特别是在材料领域。这项研究的总体假设是，基于硫醇-烯的点击化学与寡核苷酸合成的协同结合将导致形成一类全新的、功能强大的寡核苷酸结构，即硫代乙醚核酸(TNA)，这些结构能够在体内、体外和生物功能应用中与DNA相互作用，如生物检测和作为适配子模拟物。硫醇-烯点击化学与合成寡核苷酸生产的协同结合将产生功能寡核苷酸，这些功能寡核苷酸具有增强的DNA结合能力，更容易规模化生产，并且比DNA和PNA结构具有明显的化学优势。总的来说，该项目的总体目标是开发和初步表征一类全新的寡核苷酸分子，这种分子将比生物寡核苷酸(DNA和RNA)以及用于DNA杂交、生物检测和其他生物功能应用的合成PNA具有显著优势。这项工作的具体目的是(I)合成碱基功能化的TNA单体，(Ii)由这些单体形成可控的序列低聚物，并评估它们与DNA进行强选择性杂交的能力，以及(Iii)展示这些TNA序列在KRAS癌基因SNP检测中的应用。

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.