An electrochemical approach for detection of DNA methylation and assay of the methyltransferase activity

Abstract

This work develops an electrochemical approach for rapid detection of the genomic DNA methylation level, assay of methyltransferase activity, and evaluation and screening of the inhibitors of methyltransferase. This method may be a help for the discovery of anticancer drugs.

Label-free electrochemical detection of human methyltransferase from tumors

“…However, electrochemical detection schemes have typically been restricted to measurements of highly purified samples because of the increased congestion and decreased accessibility of surface (vs. solution) platforms. Electrochemistry has been used to detect nucleic acids with high sensitivity and without the need for PCR amplification in bacterial lysate and serum (18)(19)(20)(21)(22), but protein detection remains a challenge (23)(24)(25)(26). In fact, although protein detection from simple serum has been accomplished (27,28), to date no reported electrochemical systems have effectively detected active protein, of any kind, from crude cell lysate.…”

Label-free electrochemical detection of methyltransferase activity and inhibitor screening based on endonuclease HpaII and the deposition of polyaniline

“…Barton reported a multiplexed chip platform to detect both bacterial (SssI) and human (Dnmt1) methyltransferase activity (Muren and Barton, 2013). Cai reported several electrochemical method using signal amplification methods (Li et al, 2012;Liu et al, 2011). Plenty of methods based on bio enzyme are also reported (Muren and Barton, 2013;Mao et al, 2013;Ronen et al, 2014;Wei et al, 2014).…”

Highly sensitive fluorescence assay of DNA methyltransferase activity via methylation-sensitive cleavage coupled with nicking enzyme-assisted signalamplification

“…However, in addition to relatively high detection limit, the sensing platforms also suffer from time-consuming DNA conjugation on AuNPs or polymerization process. Very recently, the electrochemical and electrogenerated chemiluminescence (ECL) biosensors have been described for DNA MTase activity assay with improved detection limits even down to 0.02 U/mL level (He et al, 2011;Li et al, 2012;Liu et al, 2011;Su et al, 2012;Wu et al, 2012). In addition, a novel bioluminescence assay based on protein expression in vitro has been reported for monitoring DNA MTase activity .…”