Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 22, Pages 8050-8057Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c01401
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Funding
- National Key R&D Program of China [2019YFA0210103]
- National Natural Science Foundation of China [22074068, 21874075]
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The RESET effect was discovered to enhance the trans-cleavage activity of CRISPR/Cas12a, allowing activation even with short single-stranded DNA. This finding enriches the strategies for CRISPR/Cas12a-based sensing applications and provides potential for sensitive and specific detection of biologically important enzymes.
The trans-cleavage activity of CRISPR/Cas12a has been widely used in biosensing applications. However, the lack of exploration on the fundamental properties of CRISPR/Cas12a not only discourages further in-depth studies of the CRISPR/Cas12a system but also limits the design space of CRISPR/Cas12a-based applications. Herein, a RESET effect (random extending sequences enhance trans-cleavage activity) is discovered for the activation of CRISPR/Cas12a trans-cleavage activity. That is, a single-stranded DNA, which is too short to work as the activator, can efficiently activate CRISPR/Cas12a after being extended a random sequence from its 3 '-end, even when the random sequence folds into secondary structures. The finding of the RESET effect enriches the CRISPR/Cas12a-based sensing strategies. Based on this effect, two CRISPR/Cas12a-based biosensors are designed for the sensitive and specific detection of two biologically important enzymes.
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