1 research outputs found
High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing
DNA
hydroxymethylation (5-hmC) is a kind of new epigenetic modification,
which plays key roles in DNA demethylation, genomic reprogramming,
and the gene expression in mammals. For further exploring the functions
of 5-hmC, it is necessary to develop sensitive and selective methods
for detecting 5-hmC. Herein, we developed a novel multiplexing electrochemical
(MEC) biosensor for 5-hmC detection based on the glycosylation modification
of 5-hmC and enzymatic signal amplification. The 5-hmC was first glycosylated
by T4 β-glucosyltransferase and then oxidated by sodium periodate.
The resulting glucosyl-modified 5-hmC (5-ghmC) was incubated with
ARP-biotin and was bound to avidin-HRP. The 5-hmC can be detected
at the subnanogram level. Finally, we performed 5-hmC detection for
mouse tissue samples and cancer cell lines. The limit of detection
of the MEC biosensor is 20 times lower than that of commercial kits
based on optical meaurement. Also, the biosensor presented high detection
specificity because the chemical reaction for 5-hmC modification can
not happen at any other unhydroxymethylated nucleic acid bases. Importantly,
benefited by its multiplexing capacity, the developed MEC biosensor
showed excellent high efficiency, which was time-saving and cost less