title: A highly sensitive G-quadruplex-based luminescent switch-on probe for the detection of polymerase 3'-5' proofreading activity.
abstract: We report herein a luminescent switch-on label-free G-quadruplex-based assay for the rapid and sensitive detection of polymerase proofreading activity using a novel iridium(III) complex as a G-quadruplex-selective probe. The interaction of the iridium(III) complex with the G-quadruplex motif facilitates the highly sensitive switch-on detection of polymerase proofreading activity. Using T4 DNA polymerase (T4 pol) as a model enzyme, the assay achieved high sensitivity and selectivity for T4 pol over other tested enzymes.
authors.affiliation: Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.
authors.forename: Ka-Ho
authors.initials: KH
authors.lastname: Leung
authors.forename: Hong-Zhang
authors.initials: HZ
authors.lastname: He
authors.forename: Hai-Jing
authors.initials: HJ
authors.lastname: Zhong
authors.forename: Lihua
authors.initials: L
authors.lastname: Lu
authors.forename: Daniel Shiu-Hin
authors.initials: DS
authors.lastname: Chan
authors.forename: Dik-Lung
authors.initials: DL
authors.lastname: Ma
authors.forename: Chung-Hang
authors.initials: CH
authors.lastname: Leung
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques; D056831:Coordination Complexes / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D057075:Enzyme Assays; D005092:Exonucleases / Q000737:chemistry; D054856:G-Quadruplexes; D007495:Iridium / Q000737:chemistry; D049408:Luminescent Agents / Q000737:chemistry*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D056831:Coordination Complexes; D015342:DNA Probes; D004277:DNA, Single-Stranded; D049408:Luminescent Agents; D007495:Iridium; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases
keywords: ACN; Exo I; Exo III; G-quadruplex; Iridium(III) complex; Luminescent; PCR; Polymerase; Proofreading; T4 DNA polymerase; T4 pol; T7 Exo; T7 exonuclease; Taq DNA polymerase; Taq pol; acetonitrile; calf-thymus DNA; ctDNA; dAMP; dGMP; dNTP; deoxyadenosine monophosphate; deoxyguanosine monophosphate; deoxyribonucleoside triphosphate; double-stranded DNA; dsDNA; exonuclease I; exonuclease III; polymerase chain reaction; single-stranded DNA; ssDNA