title: High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing.
abstract: 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.
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Shixing
authors.initials: S
authors.lastname: Chen
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Yanzhi
authors.initials: Y
authors.lastname: Dou
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.|College of Sciences, Shanghai University , Shanghai 200444, China.
authors.forename: Zhihan
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Fuwu
authors.initials: F
authors.lastname: Li
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Su
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Chunhai
authors.initials: C
authors.lastname: Fan
authors.affiliation: Division of Physical Biology and Bioimaging Centre, Shanghai Synchrotron Radiation Facility, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences , Shanghai 201800, China.
authors.forename: Shiping
authors.initials: S
authors.lastname: Song
mesh_terms: D000818:Animals; D017122:Bacteriophage T4 / Q000201:enzymology; D015374:Biosensing Techniques / Q000191:economics; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D019175:DNA Methylation; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D055664:Electrochemical Techniques / Q000191:economics; D044127:Epigenesis, Genetic; D023281:Genomics; D005964:Glucosyltransferases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D057230:Limit of Detection; D051379:Mice; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry
chemical_list: C540838:5-hydroxymethyl-2'-deoxycytidine; D003841:Deoxycytidine; D010504:Periodic Acid; D004247:DNA; C009288:metaperiodate; D005964:Glucosyltransferases