Issue 1, 2024

Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

Abstract

CRISPR technology has gained widespread adoption for pathogen detection due to its exceptional sensitivity and specificity. Although recent studies have investigated the potential of high-aspect-ratio microstructures in enhancing biochemical applications, their application in CRISPR-based detection has been relatively rare. In this study, we developed a FRET-based biosensor in combination with high-aspect-ratio microstructures and Cas12a-mediated trans-cleavage for detecting HPV 16 DNA fragments. Remarkably, our results show that micropillars with higher density exhibit superior molecular binding capabilities, leading to a tenfold increase in detection sensitivity. Furthermore, we investigated the effectiveness of two surface chemical treatment methods for enhancing the developed FRET assay. A simple and effective approach was also developed to mitigate bubble generation in microfluidic devices, a crucial issue in biochemical reactions within such devices. Overall, this work introduces a novel approach using micropillars for CRISPR-based viral detection and provides valuable insights into optimizing biochemical reactions within microfluidic devices.

Graphical abstract: Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

Supplementary files

Article information

Article type
Paper
Submitted
14 sep 2023
Accepted
19 nov 2023
First published
20 nov 2023

Lab Chip, 2024,24, 47-55

Micropillar enhanced FRET-CRISPR biosensor for nucleic acid detection

M. Bao, S. J. Dollery, F. Yuqing, G. J. Tobin and K. Du, Lab Chip, 2024, 24, 47 DOI: 10.1039/D3LC00780D

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