CRISPR-Cas12a Coupled with Platinum Nanoreporter for Visual Quantification of SNVs on a Volumetric Bar-Chart Chip

Anal Chem. 2019 Oct 1;91(19):12384-12391. doi: 10.1021/acs.analchem.9b02925. Epub 2019 Sep 10.

Abstract

Methods that can detect and quantify single nucleotide variations (SNVs)/single nucleotide polymorphisms (SNPs) are greatly needed in the bioanalytical measurement of gene mutations and polymorphisms. Herein a visual and instrument-free SNV quantification platform is developed. Platinum nanoparticles tethered to magnetic beads by single-stranded DNAs are designed as quantitative readout reporters for a CRISPR-Cas12a nucleic acid detection system. The integration of platinum nanoreporter and CRISPR-Cas system with a volumetric bar-chart chip realizes the volumetric quantification of nucleic acids. This platform enables quantification of multiple cancer mutations in pure DNA samples and mock cell-free DNA samples in serum, with allelic fractions as low as 0.01%. This platform could have great potential in the quantification of SNVs/SNPs as well as other types of nucleic acid targets at the point of care.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • DNA, Single-Stranded / chemistry
  • Feasibility Studies
  • Magnets / chemistry
  • Metal Nanoparticles / chemistry*
  • Microspheres
  • Nanotechnology / instrumentation*
  • Neoplasms / genetics
  • Platinum / chemistry*
  • Polymorphism, Single Nucleotide*

Substances

  • DNA, Single-Stranded
  • Platinum