Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools (Review)

J Nanobiotechnology. 2022 Jan 21;20(1):41. doi: 10.1186/s12951-022-01246-7.

Abstract

Early detection of viral pathogens by DNA-sensors in clinical samples, contaminated foods, soil or water can dramatically improve clinical outcomes and reduce the socioeconomic impact of diseases such as COVID-19. Clustered regularly interspaced short palindromic repeat (CRISPR) and its associated protein Cas12a (previously known as CRISPR-Cpf1) technology is an innovative new-generation genomic engineering tool, also known as 'genetic scissors', that has demonstrated the accuracy and has recently been effectively applied as appropriate (E-CRISPR) DNA-sensor to detect the nucleic acid of interest. The CRISPR-Cas12a from Prevotella and Francisella 1 are guided by a short CRISPR RNA (gRNA). The unique simultaneous cis- and trans- DNA cleavage after target sequence recognition at the PAM site, sticky-end (5-7 bp) employment, and ssDNA/dsDNA hybrid cleavage strategies to manipulate the attractive nature of CRISPR-Cas12a are reviewed. DNA-sensors based on the CRISPR-Cas12a technology for rapid, robust, sensitive, inexpensive, and selective detection of virus DNA without additional sample purification, amplification, fluorescent-agent- and/or quencher-labeling are relevant and becoming increasingly important in industrial and medical applications. In addition, CRISPR-Cas12a system shows great potential in the field of E-CRISPR-based bioassay research technologies. Therefore, we are highlighting insights in this research direction.

Keywords: Bioanalytical system; COVID-19; CRISPR-Cas; CRISPR–Cas12a; DNA-biosensors; SARS-CoV-2 virus.

Publication types

  • Review

MeSH terms

  • Animals
  • Biosensing Techniques / methods
  • Biosensing Techniques / trends
  • COVID-19 / virology
  • CRISPR-Cas Systems / physiology*
  • DNA, Viral / analysis
  • DNA, Viral / isolation & purification*
  • Environmental Pollutants / analysis
  • Environmental Pollutants / isolation & purification
  • Food Contamination / analysis
  • Humans
  • Molecular Typing / methods
  • Molecular Typing / trends
  • Nucleic Acid Amplification Techniques* / methods
  • Nucleic Acid Amplification Techniques* / trends
  • SARS-CoV-2 / genetics
  • Virology / methods
  • Virology / trends
  • Virus Diseases / classification
  • Virus Diseases / diagnosis
  • Virus Diseases / virology

Substances

  • DNA, Viral
  • Environmental Pollutants