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

摘要

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.