Chemo-enzymatic synthesis of selenium-modified nucleosides, nucleotides and nucleic acids

核酸的结构研究对于理解生命过程中的大分子功能和机制至关重要。然而，核酸晶体学存在着巨大的挑战，这减慢了该领域的结构确定和深入研究。硒衍生物核酸（Selenium-derivative nucleic acids, SeNA）在核酸结构和功能研究中显示出其独特的价值，是黄教授团队为解决核酸和蛋白质-核酸复合物的相问题，促进其结晶和高分辨率而率先开展的研究。然而，化学合成SeNA往往具有挑战性，复杂，成本和劳动密集型，最终产品收率低。化学酶合成路线是一个有价值的替代，因为反应是高度特异性的，可持续的，并显示出高的最终产品收率。一系列核苷类似物以及一些核苷酸的化学酶合成已经在Neubauers教授的小组中成功证明。在中德联合合作中，将建立高效的SeNAs生产协议。通过利用双方的专业知识，含硒核苷和核苷酸将在化学酶过程中生产。RNA聚合酶或相关酶将被优化为使用硒核苷酸作为底物来高效合成SeNA。将演示使用硒修饰RNA分析HIV-Rev-RRE复合物的概念验证。RRE- rna与细胞内靶标（RRE）的相互作用将通过晶体学进行研究。所述方法的发展将有力地促进蛋白质-核酸相互作用的结晶研究。硒- na的应用不仅限于晶体学。通过这个合作研究项目，se -RNA（如反义RNA、适配体、sirna和microRNAs）将广泛应用于研究团体和生物技术和制药行业。Se-RNAs还将促进疾病分子机制的研究和治疗方法的发现。

Structural studies of nucleic acids are crucially important to understand the macromolecules function and mechanisms involved in the life process. However, great challenges exist in nucleic acid crystallography, which slow down the structure determination and depth study in this field. Selenium-derivative nucleic acids (SeNA) have shown its unique value in nucleic acid structural and functional studies, which have been pioneered by Prof. Huangs group in order to solve the phase problem and to facilitate crystallization and high resolution of nucleic acid and protein-nucleic acid complex. However, the synthesis of SeNA by chemical synthesis is often challenging, complicated, cost- and labor-intensive, which shows low final product yields. Chemo-enzymatic synthesis routes are a valuable alternative as the reactions are highly specific, sustainable and show high final product yields. The chemo-enzymatic synthesis of a series of nucleoside analogs as well as some of the nucleotides has been successfully proven in Prof. Neubauers group. In the joint Sino-German collaboration, efficient protocols for the production of SeNAs will be established. By taking advantage of the expertise of both partners, Se-containing nucleosides and nucleotides will be produced in a chemo-enzymatic process. RNA polymerases or related enzymes will be optimized to use Se-nucleotides as substrates for the efficient synthesis of SeNA. Proof-of-concept will be demonstrated using Se-modified RNA to analyze the HIV-Rev-RRE complex. The interaction of the RRE-RNA with their intracellular target (RRE) will be studied by crystallography. The development of the described method will strongly facilitate crystallization studies for protein-nucleic acid interaction. Application of Se-NA is not only limited to crystallography. Through this collaborative research project, Se-RNAs (such as antisense RNA, aptamers, siRNAs, and microRNAs) will be broadly available for the research communities and biotech & pharmaceutical industries. Se-RNAs will also facilitate disease molecular mechanism research and therapeutic discovery.