An ultrasensitive label-free RNase H assay based on in vitro transcription of fluorogenic light-up aptamer.

ABSTRACT: Herein, we proposed a label-free method to identify RNase H activity by utilizing in vitro transcription of fluorogenic light-up aptamers. In this work, we employed the specially designed two pivotal components of the hairpin substrate probe (HP) containing an RNA/DNA chimeric stem region and the template probe (TP) as a transcription template, and the RNase H activity was made to lead to the formation of a complete ds T7 promoter. T7 RNA polymerase could then promote in vitro transcription to generate numerous light-up RNA aptamers that result in significant fluorescence enhancements upon binding to the cognate fluorogenic dye. By leveraging this deliberate design principle, we identified RNase H activity ultrasensitively as low as 0.000156 U mL^-1 with excellent s

SUBMITTER: Lee J 

PROVIDER: S-EPMC10964761 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature