Rapid and Label-Free Strategy to Isolate Aptamers for Metal Ions

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• 作者：Hao Qu ; Andrew T. Csordas ; Jinpeng Wang ; Seung Soo Oh ; Michael S. Eisenstein ; Hyongsok Tom Soh
• 刊名：ACS Nano
• 出版年：2016
• 出版时间：August 23, 2016
• 年：2016
• 卷：10
• 期：8
• 页码：7558-7565
• 全文大小：427K
• 年卷期：0
• ISSN：1936-086X

文摘

Generating aptamers that bind to specific metal ions is challenging because existing aptamer discovery methods typically require chemical labels or modifications that can alter the structure and properties of the ions. In this work, we report an aptamer discovery method that enables us to generate high-quality structure-switching aptamers (SSAs) that undergo a conformational change upon binding a metal ion target, without the requirement of labels or chemical modifications. Our method is more efficient than conventional selection methods because it enables direct measurement of target binding via fluorescence-activated cell sorting (FACS), isolating only the desired aptamers with the highest affinity. Using this strategy, we obtained a highly specific DNA SSA with ∼30-fold higher affinity than the best aptamer for Hg²⁺ in the literature. We also discovered DNA aptamers that bind to Cu²⁺ with excellent affinity and specificity. Both aptamers were obtained within four rounds of screening, demonstrating the efficiency of our aptamer discovery method. Given the growing availability of FACS, we believe our method offers a general strategy for discovering high-quality aptamers for other ions and small-molecule targets in an efficient and reproducible manner.