Journal Article

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

Dronina, Julija, Samukaite-Bubniene, Urte, Ramanavicius, Arunas

2022

Overview

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. Graphical Abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adaptation / Adenoviruses / Animals / Bacteria / Bioanalytical system / Bioassays / Biosensing Techniques - methods / Biosensing Techniques - trends / Biosensors / Biotechnology / Chemistry / Chemistry and Materials Science / Classification / Cleavage / COVID-19 / COVID-19 - virology / CRISPR / CRISPR-Cas / CRISPR-Cas Systems - physiology / CRISPR–Cas12a / Cutting tools / Deoxyribonucleic acid / DNA / DNA viruses / DNA, Viral - analysis / DNA, Viral - isolation & purification / DNA-biosensors / Employment / Environmental Pollutants - analysis / Environmental Pollutants - isolation & purification / Enzymes / Fluorescence / Food contamination / Food Contamination - analysis / Genome editing / Genomes / gRNA / Health aspects / Humans / Metabolism / Molecular Medicine / Molecular Typing - methods / Molecular Typing - trends / Nanomaterials / Nanoparticles / Nanotechnology / Nucleic Acid Amplification Techniques - methods / Nucleic Acid Amplification Techniques - trends / Nucleic acids / Nucleotide sequence / Prevention / Proteins / Reagents / Review / SARS-CoV-2 - genetics / SARS-CoV-2 virus / Sediment pollution / Sensors / Soil contamination / Soil pollution / Soil water / Target recognition / Technology / Viral infections / Virology - methods / Virology - trends / Virus Diseases - classification / Virus Diseases - diagnosis / Virus Diseases - virology / Viruses