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

核酸的结构研究对于理解大分子在生命过程中的功能和机制至关重要。然而，核酸结晶学面临着巨大的挑战，阻碍了这一领域的结构确定和深入研究。硒衍生核酸(SENA)在核酸结构和功能研究中显示出其独特的价值，这是黄教授为了解决相问题，促进核酸和蛋白质-核酸复合体的结晶和高分辨率而开创的。然而，化学合成法合成SENA往往具有挑战性、复杂性、成本和劳动力密集性，最终产品收率较低。化学-酶法合成路线是一个有价值的选择，因为反应具有高度的特异性，可持续，并显示出高的最终产品产率。Neubauers教授的小组已经成功地证明了一系列核苷类似物以及一些核苷酸的化学-酶法合成。在中德联合合作中，将建立高效的SENA生产协议。通过利用双方的专业知识，将在化学-酶过程中生产含硒核苷和核苷酸。将对RNA聚合酶或相关酶进行优化，以使用Se-核苷酸作为底物高效合成SENA。概念验证将使用Se修饰的RNA来分析HIV-Rev-RRE复合体。RRE-RNA与其胞内靶点(RRE)的相互作用将通过结晶学进行研究。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。Se-Na的应用不仅限于结晶学。通过这一合作研究项目，Se-RNAs(如反义RNA、适配子、siRNAs和microRNAs)将广泛应用于研究界和生物技术和制药行业。SE-RNAs还将促进疾病分子机制研究和治疗发现。

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.