Light-Start CRISPR-Cas12a Reaction with Caged crRNA Enables Rapid and Sensitive Nucleic Acid Detection

被引：153

作者：

Hu, Menglu ^{[1
]}

Liu, Ruhan ^{[1
]}

Qiu, Zhiqiang ^{[1
]}

Cao, Feng ^{[1
]}

Tian, Tian ^{[1
]}

Lu, Yunxin ^{[4
]}

Jiang, Yongzhong ^{[5
]}

Zhou, Xiaoming ^{[1
,2
,3
]}

机构：

[1] South China Normal Univ, Sch Life Sci, Guangzhou 510631, Peoples R China

[2] South China Normal Univ, Coll Biophoton, MOE Key Lab Laser Life Sci, Guangzhou 510631, Peoples R China

[3] South China Normal Univ, Coll Biophoton, Guangdong Prov Key Lab Laser Life Sci, Guangzhou 510631, Peoples R China

[4] Sun Yat Sen Univ, Canc Ctr, Collaborat Innovat Ctr Canc Med, State Key Lab Oncol South China, Guangzhou 510060, Peoples R China

[5] Hubei Prov Ctr Dis Control & Prevent Hubei CDC, Inst Hlth Inspect & Testing, Wuhan 430079, Hubei, Peoples R China

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2023年 / 62卷 / 23期

基金：

中国国家自然科学基金;

关键词：

CRISPR-Cas12a; Caged crRNA; Nucleic Acid Detection; One-Pot Assay; Photoactivation; EPSTEIN-BARR-VIRUS; CRISPR; AMPLIFICATION; DNA; COVID-19;

D O I：

10.1002/anie.202300663

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The clustered regularly interspaced short palindromic repeats (CRISPR) system is a promising platform for nucleic acid detection. Regulating the CRISPR reaction would be extremely useful to improve the detection efficiency and speed of CRISPR diagnostic applications. Here, we have developed a light-start CRISPR-Cas12a reaction by employing caged CRISPR RNA (crRNA). When combined with recombinase polymerase amplification, a robust photocontrolled one-pot assay is achieved. The photocontrolled one-pot assay is simpler and is 50-fold more sensitive than the conventional assay. This improved detection efficiency also facilitates the development of a faster CRISPR diagnostic method. The detection of clinical samples demonstrated that 10-20 min is sufficient for effective detection, which is much faster than the current gold-standard technique PCR. We expect this advance in CRISPR diagnostics to promote its widespread detection applications in biomedicine, agriculture, and food safety.

引用

页数：8

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