Thiol-oligonucleotides for reversible chemical ligation and replication

用于可逆化学连接和复制的硫醇寡核苷酸

• 批准号: 279351736
• 负责人: Professor Dr. Günter von Kiedrowski
• 金额: --
• 依托单位: Lehrstuhl für Organische Chemie I - Bioorganische Chemie (von Kiedrowski)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/279351736?language=en
• 关键词: Thiol; oligonucleotides; reversible; chemical; ligation

Oligonucleotide templated syntheses offer a variety of applications in sequence specific programmable reactions. Chemically produced artificial oligonucleotide-systems were designed, which exist in gene diagnostics, drug screening or nano-constructs applications. Despite the wide variety of established ligation reactions of oligonucleotides, the number of reversible or switchable covalent linkage methods is limited. The project aims to contribute with a new ligation chemistry here. The ligation is based on thiol-disulfide exchange reactions that make use of the basic idea of the isosteric replacement of a 3-5-phosphodiester bond against a 3-5-disulfide bond. In this case, half-lives in the order of seconds could be achieved. The ligation is sequence selective, it can be shaped reversible or irreversible, and it proves to be sensitive to the redox state and pH-value. The project aims to exploit the full potential of the reversible disulfide linkage. A set of nucleic acid modifications is to be completed with the existent chemical synthesis methods and used in new ligation experiments. The feasibility of a directed chemical ligation, a template-directed polymerization and a dynamic DNA library is to be investigated. Furthermore, the thiol chemistry should be developed for new click-ligation reactions. The innovation of the project can be found in the configurability of the oligonucleotide ligation into reversible and irreversible direction, which can be used for the preparation of stable oligomeres out of dynamic systems. In combination with an already shown PCR amplification of disulfide oligonucleotides new applications in the context of biomedicine and DNA-nanotechnology can result.

寡核苷酸模板合成在序列特异性可编程反应中提供了各种应用。化学生产的人工寡核苷酸系统被设计出来，存在于基因诊断、药物筛选或纳米结构应用中。尽管已建立的寡核苷酸连接反应种类繁多，但可逆或可切换的共价连接方法的数量有限。该项目旨在为这里的一种新的连接化学做出贡献。该连接反应基于硫醇-二硫键交换反应，该反应利用了3-5-磷酸二酯键与3-5-二硫键的等位取代的基本思想。在这种情况下，可以实现数秒量级的半衰期。该连接具有序列选择性，可以形成可逆或不可逆的形状，并且对氧化还原状态和pH值很敏感。该项目旨在充分挖掘可逆二硫键的潜力。一组核酸修饰将用现有的化学合成方法完成，并用于新的连接实验。对定向化学连接、模板定向聚合和动态DNA文库的可行性进行了研究。此外，还应开发新的点击连接反应的硫醇化学。该项目的创新之处在于寡核苷酸连接可配置为可逆和不可逆方向，可用于从动态体系中制备稳定的低聚物。与已经展示的二硫代寡核苷酸的聚合酶链式反应扩增相结合，可以在生物医学和DNA纳米技术方面产生新的应用。