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

Author:

Hu Menglu1,Liu Ruhan1,Qiu Zhiqiang1,Cao Feng1,Tian Tian1,Lu Yunxin2,Jiang Yongzhong3,Zhou Xiaoming14ORCID

Affiliation:

1. School of Life Sciences South China Normal University Guangzhou 510631 China

2. State Key Laboratory of Oncology in South China Collaborative Innovation Center for Cancer Medicine Sun Yat-sen University Cancer Center Guangzhou 510060 China

3. Institute of Health Inspection and Testing Hubei Provincial Center for Disease Control and Prevention (Hubei CDC) Wuhan, Hubei 430079 China

4. MOE Key laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science College of Bio-photonics South China Normal University Guangzhou 510631 China

Abstract

AbstractThe 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.

Funder

National Natural Science Foundation of China

Publisher

Wiley

Subject

General Medicine

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