Light-Start CRISPR-Cas12a Reaction with Caged crRNA Enables Rapid and Sensitive Nucleic Acid Detection.
Angew Chem Int Ed Engl
; 62(23): e202300663, 2023 06 05.
Article
in English
| MEDLINE | ID: covidwho-2308962
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 10-20â
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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
CRISPR-Cas Systems
/
RNA, Guide, CRISPR-Cas Systems
Type of study:
Diagnostic study
/
Prognostic study
Language:
English
Journal:
Angew Chem Int Ed Engl
Year:
2023
Document Type:
Article
Affiliation country:
Anie.202300663
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