CRISPR-cas technology: A key approach for SARS-CoV-2 detection

Abstract

The CRISPR (Clustered Regularly Spaced Short Palindromic Repeats) system was first discovered in prokaryotes as a unique immune mechanism to clear foreign nucleic acids. It has been rapidly and extensively used in basic and applied research owing to its strong ability of gene editing, regulation and detection in eukaryotes. Hererin in this article, we reviewed the biology, mechanisms and relevance of CRISPR-Cas technology and its applications in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis. CRISPR-Cas nucleic acid detection tools include CRISPR-Cas9, CRISPR-Cas12, CRISPR-Cas13, CRISPR-Cas14, CRISPR nucleic acid amplification detection technology, and CRISPR colorimetric readout detection system. The above CRISPR technologies have been applied to the nucleic acid detection, including SARS-CoV-2 detection. Common nucleic acid detection based on CRISPR derivation technology include SHERLOCK, DETECTR, and STOPCovid. CRISPR-Cas biosensing technology has been widely applied to point-of-care testing (POCT) by targeting recognition of both DNA molecules and RNA Molecules.

Lateral Flow Assay: A Summary of Recent Progress for Improving Assay Performance

“…They are compatible with isothermal nucleic acid amplification and simple LF dipstick readout that enables sensitive/specific on-site testing [ 76 ]. Isothermal molecular amplification, such as reverse-transcription (RT)-loop-mediated isothermal amplification (LAMP) or RT-recombinase polymerase amplification (RPA), is employed as a first step to boost the assay performance, followed by a CRISPR-based recognition step triggered through sequence-specific detection of target nucleic acids using a guiding RNA (gRNA) or CRISPR RNA (crRNA)-Cas compound [ 77 , 78 ]. Cas12 or Cas13 enzymes are frequently utilized, which collaterally cleave a reporter molecule once activated by attaching to the complementary target sequence.…”

Application of CRISPR/Cas Technology in Biological Nano‐Analysis

“…At the same time, researchers are constantly improving the performance of the CRISPR/Cas system, taking advantage of its high target sequence specificity and programmability to meet the requirements of specificity, sensitivity, speed, cost and simplicity in molecular diagnostic processes. In previous reviews, there have been discussions on the regulatory mechanisms, classification, [7] biological editing in food and agriculture, [8] non nucleic acid testing, [9] and nucleic acid testing [10] of the CRISPR‐Cas system.…”

Multi-Level Biomarkers for Early Diagnosis of Ischaemic Stroke: A Systematic Review and Meta-Analysis

“…In recent years, with the advancement of point-of-care testing (POCT) technology, rapid testing has gradually become a reality for molecules ranging from DNA, RNA, proteins to metabolites, which could provide results within 5–20 min from blood sampling [ 5 , 6 , 7 ]. POCT devices are usually portable and user-friendly, leading to rapid expansion in applications across emergency departments, intensive care units, hospital wards, outpatient departments, and primary healthcare institutions [ 8 , 9 , 10 ].…”