Description

NPOM-dt was embedded into the crRNA of LbCas12a to evaluate whether Cas12a-based nucleic acid detection could be designed as a light-activated model. NPOM was modified on the heterocyclic ring of thymine and was shown to rapidly degrade under 365 nm UV light. First, the feasibility of its light-controlled activity targeting African swine fever virus (ASFV) DNA was assessed. By inserting different amounts of NPOM-dt (2 to 6) into the crRNA, the silencing effect and light-mediated activity recovery efficiency were evaluated. The crRNA activity detection was based on LbCas12a trans-cleavage, using a classical fluorescence method.

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