Label-free and sensitive detection of RNA demethylase FTO with primer generation rolling circle amplification
Xiaoxia Han, Yueying Li, Ziyue Wang, Liu L, Jian‐Ge Qiu, Bingjie Liu, Chun‐yang Zhang
Abstract
We develop for the first time a label-free fluorescent method for sensitive detection of fat mass and obesity-associated protein (FTO) activity using MazF-mediated primer generation rolling circle amplification. This method is very simple with ultrahigh sensitivity and good specificity, and it can detect FTO activity at the single-cell level. Moreover, this method can be applied for the measurement of kinetic parameters and the screening of FTO inhibitors.
Topics & Concepts
FTO genePrimer (cosmetics)Rolling circle replicationRNAFluorescenceChemistrySensitivity (control systems)Molecular biologyBiologyEnzymeBiochemistryGenePhysicsPolymeraseQuantum mechanicsElectronic engineeringEngineeringGenotypeOrganic chemistryPolymorphism (computer science)RNA modifications and cancerRNA Research and SplicingRNA and protein synthesis mechanisms