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

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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.