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High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing
Chen, SX; Dou, YZ; Zhao, ZH; Li, FW; Su, J; Fan, CH; Song, SP; Song, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai 201800, Peoples R China.; Song, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Bioimaging Ctr, Shanghai Synchrotron Radiat Facil,CAS Key Lab Int, Shanghai 201800, Peoples R China.
2016
Source PublicationANALYTICAL CHEMISTRY
ISSN0003-2700
Volume88Issue:7Pages:3476—3480
Subtype期刊文献
AbstractDNA 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 beta-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.
KeywordPrinted Amperometric Biosensor Mammalian Dna Tet Proteins 5-hydroxymethylcytosine Methylation 5-methylcytosine Brain 5-carboxylcytosine Quantification Cancer
DOI10.1021/acs.analchem.6b00230
Indexed BySCI
Language英语
WOS IDWOS:000373656300011
Citation statistics
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/25746
Collection中科院上海应用物理研究所2011-2018年
Corresponding AuthorFan, CH; Song, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai 201800, Peoples R China.; Song, SP (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Bioimaging Ctr, Shanghai Synchrotron Radiat Facil,CAS Key Lab Int, Shanghai 201800, Peoples R China.
Recommended Citation
GB/T 7714
Chen, SX,Dou, YZ,Zhao, ZH,et al. High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing[J]. ANALYTICAL CHEMISTRY,2016,88(7):3476—3480.
APA Chen, SX.,Dou, YZ.,Zhao, ZH.,Li, FW.,Su, J.,...&Song, SP .(2016).High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing.ANALYTICAL CHEMISTRY,88(7),3476—3480.
MLA Chen, SX,et al."High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing".ANALYTICAL CHEMISTRY 88.7(2016):3476—3480.
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