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.
MeSH terms
-
Alpha-Ketoglutarate-Dependent Dioxygenase FTO / analysis*
-
Alpha-Ketoglutarate-Dependent Dioxygenase FTO / metabolism
-
Cell Line, Tumor
-
DNA, Single-Stranded / chemistry
-
DNA, Single-Stranded / metabolism*
-
DNA-Binding Proteins / metabolism
-
Endoribonucleases / metabolism
-
Escherichia coli Proteins / metabolism
-
Humans
-
Limit of Detection
-
Native Polyacrylamide Gel Electrophoresis
-
Nucleic Acid Amplification Techniques / methods*
-
Single-Cell Analysis
-
Spectrometry, Fluorescence
Substances
-
DNA, Single-Stranded
-
DNA-Binding Proteins
-
Escherichia coli Proteins
-
MazF protein, E coli
-
Alpha-Ketoglutarate-Dependent Dioxygenase FTO
-
Endoribonucleases