期刊
BIOSENSORS & BIOELECTRONICS
卷 64, 期 -, 页码 449-455出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2014.09.044
关键词
DNA methylation; Methyltransferase; Photoelectrochemistry; Exciton energy transfer; Enzyme cleavage
类别
资金
- National Natural Science Foundation of China [21105050, 21475057]
- Program for New Century Excellent Talents in University [NCET-12-0256]
- Ministry of Education of the People's Republic of China [IRT1148]
- Research Found for the Doctoral Program of Higher Education of China [20113223120004]
- Foundation of the Jiangsu Education Committee [14KJB150015]
- Priority Academic Program Development of the Jiangsu Higher Education Institutions
- Deanship of Scientific Research at King Saud University [RGP-VPP-029]
Highly sensitive DNA methyltransferase (MTase) activity and inhibitor screening photoelectrochemical (PEC) assay was developed based on the exciton energy transfer (EET) effect coupled with site-specific cleavage of restriction endonuclease (HpaII). The assay was designed by integrating the Au nanoparticles (NPs) labeled probe DNA (pDNA-Au) with CdSe quantum dots (QDs). The strong EET effect between Au NPs and CdSe QDs resulted in the dramatic decrease of photocurrent signal. The pDNA carried a sensing region for specifically recognizing target DNA (tDNA) and hybridizing with it to form a DNA duplex. With the site-specific cleavage of HpaIl, the DNA duplex could be cleaved and Au NPs would be released, which broke the EET and resulted in the restoration of photocurrent signal. However, when the DNA duplex was methylated by M.SssI MTase, this cleavage of HpaII was blocked, and therefore the unbroken BET effect kept the lower photocurrent signal. That was, the restored photocurrent was inversely proportional to the MTase activity. Based on this strategy, the PEC assay could determine as low as similar to 0.0042 U/mL of M. SssI MTase with a linear range from 0.01 to 150 U/mL. In addition, the assay could be used for the screening of the inhibitors of MTase. This PEC assay provides a promising platform for monitoring the activity and inhibition of DNA MTase, and thus shows a great potential in cancer diagnostics and anticancer drugs discovery. (C) 2014 Elsevier B.V. All rights reserved.
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