Sau Sujay P, Fahmi Nour Eddine, Liao Jen-Yu, Bala Saikat, Chaput John C
Department of Pharmaceutical Sciences, University of California , Irvine, California 92697, United States.
The Biodesign Institute, Arizona State University , Tempe, Arizona 85287-5301, United States.
J Org Chem. 2016 Mar 18;81(6):2302-7. doi: 10.1021/acs.joc.5b02768. Epub 2016 Feb 26.
Recent advances in polymerase engineering have made it possible to copy information back and forth between DNA and artificial genetic polymers composed of TNA (α-L-threofuranosyl-(3',2') nucleic acid). This property, coupled with enhanced nuclease stability relative to natural DNA and RNA, warrants further investigation into the structural and functional properties of TNA as an artificial genetic polymer for synthetic biology. Here, we report a highly optimized chemical synthesis protocol for constructing multigram quantities of TNA nucleosides that can be readily converted to nucleoside 2'-phosphoramidites or 3'-triphosphates for solid-phase and polymerase-mediated synthesis, respectively. The synthetic protocol involves 10 chemical transformations with three crystallization steps and a single chromatographic purification, which results in an overall yield of 16-23% depending on the identity of the nucleoside (A, C, G, T).
聚合酶工程学的最新进展使得在DNA与由TNA（α-L-苏糖呋喃糖基-(3',2')核酸）组成的人工遗传聚合物之间来回复制信息成为可能。这一特性，再加上相对于天然DNA和RNA而言增强的核酸酶稳定性，使得有必要进一步研究TNA作为合成生物学中人工遗传聚合物的结构和功能特性。在此，我们报告了一种高度优化的化学合成方案，用于构建数克量的TNA核苷，这些核苷可分别轻松转化为核苷2'-亚磷酰胺或3'-三磷酸，用于固相合成和聚合酶介导的合成。该合成方案涉及10步化学转化、三个结晶步骤和一次柱色谱纯化，根据核苷（A、C、G、T）的种类不同，总产率为16%-23%。
