Department of Chemistry
Detection of 3′ → 5′ exonuclease activity using a metal-based luminescent switch-on probe
A luminescent iridium(III) complex has been discovered to be selective for G-quadruplex DNA, and was employed in a label-free G-quadruplex-based detection assay for 3'→5' exonuclease activity in aqueous solution. A proof-of-concept of this assay has been demonstrated by using prokaryotic exonuclease III (ExoIII) as a model enzyme. In this assay, a G-quadruplex-forming hairpin oligonucleotide (hairpin-G4 DNA, 5'-GAG3TG4AG3TG4A2GCAGA2G2ATA2CT2C4AC3TC4AC3TC-3') initially exists in a duplex conformation, resulting in a low luminescence signal due to the weak interaction between the iridium(III) complex and duplex DNA. Upon digestion by ExoIII, the guanine-rich sequence is released and folds into a G-quadruplex, which greatly enhances the luminescence emission of the iridium(III) probe. This method was highly sensitive for 3'→5' exonuclease over other DNA-modifying enzymes. © 2013 Elsevier Inc.
Exonuclease, G-quadruplex, Iridium, Luminescence, Metal complex
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He, H., Chan, W., Mak, T., Liu, L., Wang, M., Chan, D., Ma, D., & Leung, C. (2013). Detection of 3′ → 5′ exonuclease activity using a metal-based luminescent switch-on probe. Methods, 64 (3), 218-223. https://doi.org/10.1016/j.ymeth.2013.08.011