Amplified Nucleic Acid Sensing Using Programmed Self-Cleaving DNAzyme

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Abstract
A newly designed target-assisted self-cleavage (TASC) probe composed of a target-binding site and a DNAzyme domain undergoes TASC when activated via hybridization with a target DNA/RNA. This self-splicing or self-dissociation reaction occurs in a catalytic manner with the probe as a substrate and the target as a catalyst, since the fragmented products are automatically released from the target, thus amplifying the sequence information of the latter under non-PCR, i.e., isothermal and enzyme/reagent-free, conditions. A fluorescence-reporting TASC probe having a fluorescein/dabsyl FRET pair across the cleavage site allows a mix-and-read discrimination of single-nucleotide differences in the target.

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