Development and application of sensitive, specific and rapid CRISPR-Cas13-based diagnosis.
2021
Nucleic acid detection is a necessary part of medical treatment and field work. However, the current detection technologies are far from ideal. A lack of timely and accessible testing for identifying cases and close contacts has allowed SARS-CoV-2, the causative virus of the ongoing COVID-19 pandemic, to spread uncontrollably. The slow and expensive detection of mutations - predictors for chronic diseases such as cancer - form a barrier to personalized treatment. A recently developed diagnostic assay is ideal and field-ready - it relies on CRISPR-Cas13. CRISPR-Cas13 works similarly to other CRISPR systems: Cas13 is guided by a crRNA to cleave next to a specific RNA target sequence. Additionally, Cas13 boasts a unique collateral cleavage activity; collateral cleavage of a fluorescent reporter detects the presence of the target sequence in sample RNA. This system forms the basis of CRISPR-Cas13 diagnostic assays. CRISPR-Cas13 assays have >95% sensitivity and >99% specificity. Detection is rapid (2 hours), inexpensive ($0.05), and portable - a test using a lateral flow strips is akin to a pregnancy test. The recent adaptation of micro-well chips facilitates high-level multiplexing and is high-throughput. In this review, we cover the development of CRISPR-Cas13 assays for medical diagnosis, discuss the advantages of CRISPR-Cas13-based diagnosis over traditional RT-PCR, and present examples of detection from real patient samples. This article is protected by copyright. All rights reserved.
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