发表刊物：Anal Chem.
摘要：Aptamers are single-stranded DNA or RNA molecules that bind specific ligands with high affinity. DNA aptamers are especially promising due to their enhanced stability and cost-effectiveness, yet their practical use often necessitates coupling with an expression platform to transduce ligand-binding events into detectable signals. To overcome this limitation, we developed Expression-SELEX, an efficient selection method that integrates a DNAzyme-based expression platform directly into the screening process. Previously, we employed this approach to isolate a phenylalanine-responsive allosteric DNAzyme with significant ligand-dependent self-cleavage activity. However, the broader applicability of Expression-SELEX for discovering diverse ligand-dependent DNAzymes and their utility in clinical detection remained unexplored. In this study, we applied Expression-SELEX to identify theophylline-responsive allosteric DNAzymes as functional replacements for traditional RNA aptamers. Our selection yielded 12,880 unique candidates, with all top 10 enriched candidates displaying both significant theophylline-dependent cleavage activity and excellent specificity, allowing effective discrimination between theophylline and caffeine. Although the cleavage products of these ligand-responsive DNAzymes could be detected through PAGE gel analysis as basic biosensors, we enhanced their functionality by incorporating a fluorophore-quencher pair for liquid-phase assays in 96-well plates. The optimized biosensors exhibited excellent performance in reaction buffer containing 1%-5% urine. These findings highlight the versatility of Expression-SELEX for rapidly generating ligand-responsive DNAzymes and biosensors, underscoring their potential for diagnostic and biomedical applications.
备注：Chen B, Lin Y, Chen H, Li H, Huang Y, Zhang S, Zhang X, Li S. Selection of Allosteric DNAzymes for Theophylline Sensing by Expression-SELEX. Anal Chem. 2025 Dec 23;97(50):27734-27744. doi: 10.1021/acs.analchem.5c04622.
论文类型：期刊论文
卷号：97
期号：50
页面范围：27734-27744
是否译文：否
发表时间：2025-12-23
收录刊物：SCI
影响因子：6.7