Abstract Nucleic acid detection is a necessary part of medical treatment and fieldwork. However, the current detection technologies are far from ideal. A lack of timely and accessible testing for identifying cases and close contacts has allowed severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2), the causative virus of the ongoing coronavirus disease‐2019 (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 h), inexpensive ($0.05 per test), and portable—a test using 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 the traditional reverse transcription polymerase chain reaction (RT‐PCR), and present examples of detection from real patient samples. Highlights In the CRISPR‐Cas13 system, Cas13 is guided by a crRNA to cleave sample RNA at a specific target sequence. Cas13 has collateral cleavage activity that when used with a fluorescent reporter detects a specific target sequence. CRISPR‐Cas13‐based diagnostics are highly sensitive (>95%), specific (>99%), rapid (<2 h), inexpensive ($0.05 per test), and portable‐recent adaptions of micro‐well chips facilitate high‐evel multiplexing and are high‐throughput. CRISPR‐Cas13‐based diagnostics have been validated in the detection of viral RNA and mutations in patient samples, including SARS‐CoV‐2 RNA during the ongoing COVID‐19 pandemic. CRISPR‐Cas13 is also applied to RNA editing, thereby creating a full system of diagnostics and therapeutics.
【저자키워드】 Coronavirus disease 2019, severe acute respiratory syndrome coronavirus‐2, Nucleic acid detection, clustered regularly interspaced short palindromic repeats (CRISPR)‐associated genes 13, specific high‐sensitivity enzymatic reporter unlocking, 【초록키워드】 Treatment, coronavirus, pandemic, Mutation, diagnostic, Diagnosis, detection, virus, diagnostics, chronic disease, COVID‐19, RNA, SARS‐CoV‐2, Spread, CRISPR, lateral flow strip, sensitivity, specificity, Patient, cleavage, reverse transcription, Viral RNA, reporter, close contact, pregnancy test, acute respiratory syndrome, sequence, fluorescent, RT‐PCR, polymerase chain, detect, lack, example, assays, form, applied, facilitate, unique, cleave, creating, 【제목키워드】 Diagnosis, development,