title: A transformer of molecular beacon for sensitive and real-time detection of phosphatases with effective inhibition of the false positive signals.
abstract: Molecular beacons (MBs) have shown great potential in measurement of enzyme activities. However, currently available methods for monitoring of phosphatases only use MBs as a signal reporter. Extra substrates for the phosphatases are needed to hybridize to the MB either as a primer or as a template. Moreover, few MB-based methods have been used to detect enzyme activities in real biological samples due to insufficient sensitivity or false positive interference signals caused by nonspecific nucleases. In this work, a novel type of fluorescent probe was designed and synthesized for monitoring of phosphatases by integrating the DNA substrate and the signaling structures into a single molecule. Such a new design not only significantly simplified the probing system and greatly enhanced the sensitivity, but also offered a practical way to guard against the false-positive signal problems in the application to real samples. The unique design of the assay format should be widely applicable to many other enzymatic assays using oligonucleotide fluorescent probes.
authors.affiliation: Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.forename: Xin
authors.initials: X
authors.lastname: Su
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liang
authors.forename: Xiaocui
authors.initials: X
authors.lastname: Zhu
authors.forename: Chen
authors.initials: C
authors.lastname: Song
authors.forename: Meiping
authors.initials: M
authors.lastname: Zhao
mesh_terms: D002138:Calibration; D005189:False Positive Reactions; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D010744:Phosphoric Monoester Hydrolases