Center for Multifunctional Biomolecular Drug Design (CEMBID) at the, Interdisciplinary Nanoscience Center (iNANO) and the Department of Chemistry, Aarhus University, 8000, Aarhus C, Denmark.
Chemistry. 2019 Sep 20;25(53):12303-12307. doi: 10.1002/chem.201903496. Epub 2019 Aug 29.
Triplex forming oligonucleotides are used as a tool for gene regulation and in DNA nanotechnology. By incorporating artificial nucleic acids, target affinity and biological stability superior to that of natural DNA may be obtained. This work demonstrates how a chimeric clamp consisting of acyclic (L)-threoninol nucleic acid (aTNA) and DNA can bind DNA and RNA by the formation of a highly stable triplex structure. The (L)-aTNA clamp is released from the target again by the addition of a releasing strand in a strand displacement type of reaction. It is shown that the clamp efficiently inhibits Bsu and T7 RNA polymerase activity and that polymerase activity is reactivated by displacing the clamp. The clamp was successfully applied to the regulation of luciferase expression by reversible binding to the mRNA. When targeting a sequence in the double stranded plasmid, 40 % downregulation of protein expression is achieved.
三链形成寡核苷酸被用作基因调控的工具和在 DNA 纳米技术中。通过引入人工核酸，可以获得优于天然 DNA 的靶亲和力和生物稳定性。这项工作展示了如何通过形成高度稳定的三链结构，由无环（L）-苏氨酸核酸（aTNA）和 DNA 组成的嵌合夹来结合 DNA 和 RNA。通过添加释放链，在链置换类型的反应中，（L）-aTNA 夹再次从靶标中释放。结果表明，夹有效地抑制了 Bsu 和 T7 RNA 聚合酶的活性，并且通过置换夹可以使聚合酶活性重新激活。该夹通过可逆结合 mRNA 成功应用于对荧光素酶表达的调控。当靶向双链质粒中的序列时，可实现 40%的蛋白表达下调。
