Label-free profiling of DNA aptamer-small molecule binding using T5 exonuclease.

ABSTRACT: In vitro aptamer isolation methods can yield hundreds of potential candidates, but selecting the optimal aptamer for a given application is challenging and laborious. Existing aptamer characterization methods either entail low-throughput analysis with sophisticated instrumentation, or offer the potential for higher throughput at the cost of providing a relatively increased risk of false-positive or -negative results. Here, we describe a novel method for accurately and sensitively evaluating the binding between DNA aptamers and small-molecule ligands in a high-throughput format without any aptamer engineering or labeling requirements. This approach is based on our new finding that ligand binding inhibits aptamer digestion by T5 exonuclease, where the extent of this inhibition correlates clo

SUBMITTER: Alkhamis O 

PROVIDER: S-EPMC7672472 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature