Universal and Naked-Eye Gene Detection Platform Based on the Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a/13a System

Gold-nanoparticles-based colorimetric assay is an attractive detection format, but is limited by the tedious and ineffective posthybridization manipulations for genomic analysis. Here, we present a new design for a colorimetric gene-sensing platform based on the clustered regularly interspaced short...

Full description

Saved in:
Bibliographic Details
Published inAnalytical chemistry (Washington) Vol. 92; no. 5; pp. 4029 - 4037
Main Authors Yuan, Chaoqun, Tian, Tian, Sun, Jian, Hu, Menglu, Wang, Xusheng, Xiong, Erhu, Cheng, Meng, Bao, Yijuan, Lin, Wei, Jiang, Jieming, Yang, Chengwei, Chen, Qian, Zhang, Huang, Wang, Heng, Wang, Xiran, Deng, Xianbo, Liao, Xiaoping, Liu, Yahong, Wang, Zhang, Zhang, Guihong, Zhou, Xiaoming
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 03.03.2020
Subjects
Online AccessGet full text

Cover

Loading…
More Information
Summary:Gold-nanoparticles-based colorimetric assay is an attractive detection format, but is limited by the tedious and ineffective posthybridization manipulations for genomic analysis. Here, we present a new design for a colorimetric gene-sensing platform based on the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system. In this strategy, programmable recognition of DNA by Cas12a/crRNA and RNA by Cas13a/crRNA with a complementary target activates the trans-ssDNA or -ssRNA cleavage. Target-induced trans-ssDNA or ssRNA cleavage triggers an aggregation behavior change for the designed AuNPs–DNA probes pair, enabling the completion of naked-eye gene detection (transgenic rice, African swine fever virus, and miRNAs as the models) within 1 h. This platform is also showing promise as a fast and inexpensive tool for bacteria identification using 16S rDNA or 16S rRNA. A CRISPR/Cas-based colorimetric platform shows superior characteristics, such as probe universality, compatibility with isothermal reaction conditions, on-site detection capability, and high sensitivity, thus, demonstrating its use as a robust next-generation gene detection platform.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.9b05597