Department of Chemistry, University of Georgia , 140 Cedar Street, Athens, Georgia 30602, United States.
Department Chemistry, York University , 4700 Keele Street, Toronto, ON M3J 1P3, Canada.
J Am Chem Soc. 2017 Oct 11;139(40):13977-13980. doi: 10.1021/jacs.7b07241. Epub 2017 Sep 27.
We describe the application of T4 DNA ligase-catalyzed DNA templated oligonucleotide polymerization toward the evolution of a diversely functionalized nucleic acid aptamer for human α-thrombin. Using a 256-membered ANNNN comonomer library comprising 16 sublibraries modified with different functional groups, a highly functionalized aptamer for thrombin was raised with a dissociation constant of 1.6 nM. The aptamer was found to be selective for thrombin and required the modifications for binding affinity. This study demonstrates the most differentially functionalized nucleic acid aptamer discovered by in vitro selection and should enable the future exploration of functional group dependence during the evolution of nucleic acid polymer activity.
我们描述了 T4 DNA 连接酶催化的 DNA 模板寡核苷酸聚合在人类 α-凝血酶多样化功能化核酸适体进化中的应用。使用包含 16 个子库的 256 成员的 ANNNN 共聚单体库，其中不同的功能基团进行了修饰，得到了一种对凝血酶具有高亲和力的高度功能化适体，解离常数为 1.6 nM。该适体被发现对凝血酶具有选择性，并且需要修饰才能具有结合亲和力。这项研究展示了通过体外选择发现的最具差异化功能化的核酸适体，应该能够促进未来在核酸聚合物活性进化过程中对功能基团依赖性的探索。
