Department of Chemistry, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
J Am Chem Soc. 2017 Mar 1;139(8):2892-2895. doi: 10.1021/jacs.6b13132. Epub 2017 Feb 20.
RNA or single-stranded DNA aptamers with 2'-F pyrimidines have been pursued to increase resistance to nucleases, and while it seems likely that these and other modifications, including the modification of purines, could be used to optimize additional properties, this has been much less explored because such aptamers are challenging to discover. Using a thermostable DNA polymerase, SFM4-3, which was previously evolved to accept nucleotides with 2'-modifications, we now report the selection of 2'-F purine aptamers that bind human neutrophil elastase (HNE). Two aptamers were identified, 2fHNE-1 and 2fHNE-2, that bind HNE with reasonable affinity. Interestingly, the 2'-F substituents facilitate the selection of specific interactions with HNE and overcome nonspecific electrostatic interactions that can otherwise dominate. The data demonstrate that inclusion of only a few 2'-F substituents can optimize properties far beyond simple nuclease resistance and that SFM4-3 should prove valuable for the further exploration and production of aptamers with properties optimized for various applications.
已经有人研究了具有 2'-F 嘧啶的 RNA 或单链 DNA 适体，以提高对核酸酶的抗性。虽然这些和其他修饰（包括嘌呤的修饰）似乎可以用于优化其他特性，但这方面的研究要少得多，因为这些适体的发现具有挑战性。使用以前经过进化以接受具有 2'-修饰的核苷酸的热稳定 DNA 聚合酶 SFM4-3，我们现在报告了选择与人中性粒细胞弹性蛋白酶（HNE）结合的 2'-F 嘌呤适体。鉴定出两种适体，2fHNE-1 和 2fHNE-2，它们与 HNE 的结合具有合理的亲和力。有趣的是，2'-F 取代基有助于选择与 HNE 的特异性相互作用，并克服否则可能占主导地位的非特异性静电相互作用。这些数据表明，仅包含少数 2'-F 取代基就可以极大地优化性能，远远超过简单的抗核酸酶性，并且 SFM4-3 应该对进一步探索和生产针对各种应用优化的特性的适体非常有价值。
