A highly sensitive and selective fluoride sensor based on a riboswitch-regulated transcription coupled with CRISPR-Cas13a tandem reaction

Yuan Ma, Quanbing Mou, Peng Yan, Zhenglin Yang, Ying Xiong, Deyue Yan, Chuan Zhang, Xinyuan Zhu, Yi Lu

Research output: Contribution to journalArticlepeer-review

Abstract

Nucleic acid sensors have realized much success in detecting positively charged and neutral molecules, but have rarely been applied for measuring negatively charged molecules, such as fluoride, even though an effective sensor is needed to promote dental health while preventing osteofluorosis and other diseases. To address this issue, we herein report a quantitative fluoride sensor with a portable fluorometer readout based on fluoride riboswitch-regulated transcription coupled with CRISPR-Cas13-based signal amplification. This tandem sensor utilizes the fluoride riboswitch to regulatein vitrotranscription and generate full-length transcribed RNA that can be recognized by CRISPR-Cas13a, triggering the collateral cleavage of the fluorophore-quencher labeled RNA probe and generating a fluorescence signal output. This tandem sensor can quantitatively detect fluoride at ambient temperature in aqueous solution with high sensitivity (limit of detection (LOD) ≈ 1.7 μM), high selectivity against other common anions, a wide dynamic range (0-800 μM) and a short sample-to-answer time (30 min). This work expands the application of nucleic acid sensors to negatively charged targets and demonstrates their potential for the on-site and real-time detection of fluoride in environmental monitoring and point-of-care diagnostics.

Original languageEnglish (US)
Pages (from-to)11740-11747
Number of pages8
JournalChemical Science
Volume12
Issue number35
DOIs
StatePublished - Sep 21 2021

ASJC Scopus subject areas

  • Chemistry(all)

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