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

Authors :
Hu, Menglu
Liu, Ruhan
Qiu, Zhiqiang
Cao, Feng
Tian, Tian
Lu, Yunxin
Jiang, Yongzhong
Zhou, Xiaoming
Source :
Angewandte Chemie; 6/5/2023, Vol. 135 Issue 23, p1-8, 8p
Publication Year :
2023

Abstract

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. [ABSTRACT FROM AUTHOR]

Subjects

Subjects :
CRISPRS
POLYMERASES
NUCLEIC acids

Details

Language :
English
ISSN :
00448249
Volume :
135
Issue :
23
Database :
Complementary Index
Journal :
Angewandte Chemie
Publication Type :
Academic Journal
Accession number :
163911117
Full Text :
https://doi.org/10.1002/ange.202300663