Fig. 1. Schematic of the rRCA–Cas reaction. (IMAGE)

Nanjing Agricultural University The Academy of Science

Fig. 1. Schematic of the rRCA–Cas reaction.

Caption

After hybridizing to the precircularized circular probe, the target miRNA is amplified into a long ssDNA product by RCA with massive trigger sequences repeats. The trigger sequence is designed as the activator of Cas12a for showing trans-cleavage activity. Then, the trigger chain is recognized by Cas12a–crRNA duplex and hybridizes to it. Cas12a was then triggered to demonstrate trans-cleavage activity, F-Q probe (an ssDNA probe labeled with a fluorophore at the 5′ end and a quencher at the 3′ end) was cleaved, resulting in separation of the quencher from the fluorophore, generating the fluorescence signal.

Credit

BioDesign Research

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