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Related Experiment Video

Updated: Aug 27, 2025

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Accelerated CRISPR/Cas12a-based small molecule detection using bivalent aptamer.

Xiuping Li1, Xiujin Chen2, Minxin Mao1

  • 1State Key Laboratory of Food Science and Technology, Jiangnan University, Lihu Road 1800, Wuxi, 214122, PR China; School of Food Science and Technology, Jiangnan University, Lihu Road 1800, Wuxi, 214122, PR China.

Biosensors & Bioelectronics
|September 30, 2022
PubMed
Summary

This study introduces a novel CRISPR/Cas biosensing method using bivalent aptamers for rapid small molecule detection. The BA-CASLFA platform enables visual, on-site detection of targets like ATP and kanamycin within minutes.

Keywords:
Adenosine 5′-triphosphateAptamer conformationCRISPR/Cas12a systemKanamycinLateral flow assay

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biosensing

Background:

  • CRISPR/Cas systems are powerful tools for nucleic acid detection.
  • Current CRISPR/Cas applications are limited to nucleic acid targets.
  • There is a need for CRISPR-based sensors capable of detecting small molecules.

Purpose of the Study:

  • To broaden the target range of CRISPR/Cas systems to small molecules.
  • To develop a rapid and visual biosensing platform for small molecules.
  • To demonstrate the versatility of the developed platform for various analytes.

Main Methods:

  • Integration of bivalent aptamers with CRISPR/Cas systems for small molecule recognition.
  • Development of a CRISPR/Cas-based lateral flow assay (BA-CASLFA) for visual readout.
  • Selection and design of bivalent aptamers for specific small molecules like ATP and kanamycin.

Main Results:

  • Bivalent aptamer significantly accelerated ATP binding and detection time.
  • The BA-CASLFA platform provided a visual "TURN ON" colorimetric signal for ATP within 26 minutes.
  • The platform demonstrated versatility by successfully detecting kanamycin.

Conclusions:

  • The developed BA-CASLFA strategy offers a generalizable approach for CRISPR-based small molecule sensing.
  • This method enables rapid, on-site detection suitable for clinical diagnosis, food safety, and environmental monitoring.
  • The integration of bivalent aptamers expands the utility of CRISPR/Cas technology beyond nucleic acids.