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

Anal Chem. 2020 Mar 3;92(5):4029-4037. doi: 10.1021/acs.analchem.9b05597. Epub 2020 Feb 17.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • African Swine Fever Virus / genetics
  • Animals
  • Bacteria / genetics
  • CRISPR-Cas Systems / genetics*
  • Colorimetry / methods*
  • DNA Probes / chemistry
  • DNA, Viral / analysis
  • DNA, Viral / chemistry
  • Gold / chemistry
  • Metal Nanoparticles / chemistry
  • MicroRNAs / analysis
  • MicroRNAs / chemistry
  • Promoter Regions, Genetic
  • RNA, Ribosomal, 16S / analysis*
  • RNA, Ribosomal, 16S / chemistry
  • Swine

Substances

  • DNA Probes
  • DNA, Viral
  • MicroRNAs
  • RNA, Ribosomal, 16S
  • Gold