Digital CRISPR systems for the next generation of nucleic acid quantification

Anthony J. Politza, Reza Nouri, Weihua Guan

Research output: Contribution to journalReview articlepeer-review

9 Scopus citations

Abstract

Digital CRISPR (dCRISPR) assays are an emerging platform of molecular diagnostics. Digital platforms introduce absolute quantification and increased sensitivity to bulk CRISPR assays. With ultra-specific targeting, isothermal operation, and rapid detection, dCRISPR systems are well-prepared to lead the field of molecular diagnostics. Here we summarized the common Cas proteins used in CRISPR detection assays. The methods of digital detection and critical performance factors are examined. We formed three strategies to frame the landscape of dCRISPR systems: (1) amplification free, (2) in-partition amplification, and (3) two-stage amplification. We also compared the performance of all systems through the limit of detection (LOD), testing time, and figure of merit (FOM). This work summarizes the details of digital CRISPR platforms to guide future development. We envision that improvements to LOD and dynamic range will position dCRISPR as the leading platform for the next generation of molecular biosensing.

Original languageEnglish (US)
Article number116917
JournalTrAC - Trends in Analytical Chemistry
Volume159
DOIs
StatePublished - Feb 2023

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Spectroscopy

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