Small-Molecule-Dependent Split Aptamer Ligation

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• 作者：Ashwani K. Sharma ; Jennifer M. Heemstra
• 刊名：Journal of the American Chemical Society
• 出版年：2011
• 出版时间：August 17, 2011
• 年：2011
• 卷：133
• 期：32
• 页码：12426-12429
• 全文大小：738K
• 年卷期：v.133,no.32(August 17, 2011)
• ISSN：1520-5126

文摘

Here we describe the first use of small-molecule binding to direct a chemical reaction between two nucleic acid strands. The reported reaction is a ligation between two fragments of a DNA split aptamer using strain-promoted azide鈥揳lkyne cycloaddition. Utilizing the split aptamer for cocaine, we demonstrate small-molecule-dependent ligation that is dose-dependent over a wide range of cocaine concentrations and is compatible with complex biological fluids such as human blood serum. Moreover, studies of split aptamer ligation at varying salt concentrations and using structurally similar analogues of cocaine have revealed new insight into the assembly and small-molecule binding properties of the cocaine split aptamer. The ability to translate the presence of a small-molecule target into the output of DNA ligation is anticipated to enable the development of new, broadly applicable small-molecule detection assays.