Amplified nucleic acid sensing using programmed self-cleaving DNAzyme.
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.
Sando
Shinsuke
S
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan. ssando@sbchem.kyoto-u.ac.jp
Sasaki
Toshinori
T
Kanatani
Keiichiro
K
Aoyama
Yasuhiro
Y
D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000235:genetics; D021881:DNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D016762:Genes, fos; D007700:Kinetics; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization / Q000379:methods; D009711:Nucleotides / Q000032:analysis* / Q000235:genetics; D012313:RNA / Q000032:analysis* / Q000235:genetics
D021881:DNA, Catalytic; D009711:Nucleotides; D012313:RNA; D004247:DNA