Simple and cost-effective fluorescent labeling of 5-hydroxymethylcytosine. (7th October 2016)

• Record Type: Journal Article
• Authors: Shahal, Tamar; Green, Ori; Hananel, Uri; Michaeli, Yael; Shabat, Doron; Ebenstein, Yuval
• Is Part Of: Methods and applications in fluorescence. Volume 4:Number 4(2016:Dec.)
• Publication Date: 2016-10-07
• Subjects: glucose oxidation -- aminooxy-Cy5 -- epigenetics -- DNA-labeling -- aldehyde reactive probes -- 5-hydroxymethylcytosine; Fluorescence spectroscopy -- Periodicals; Fluorescence -- Periodicals; Spectrometry, Fluorescence -- Periodicals; Fluorescence -- Periodicals; Fluorescence; Fluorescence spectroscopy; Periodicals; 535.352
• DOI: 10.1088/2050-6120/4/4/044003
• Languages: English
• ISSNs: 2050-6120

Abstract

Abstract: The nucleobase 5-hydroxymethylcytosine (5-hmC), a modified form of cytosine, is an important epigenetic mark related to regulation of gene expression. 5-hmC levels are highly dynamic during early development and are modulated during the progression of neurodegenerative disease and cancer. We describe a spectroscopic method for the global quantification of 5-hmC in genomic DNA. This method relies on the enzymatic glucosylation of 5-hmC, followed by a glucose oxidation step that results in the formation of aldehyde moieties that are covalently linked to a fluorescent reporter by oxime ligation. The fluorescence intensity of the labeled sample is directly proportional to its 5-hmC content. We show that this simple and cost-effective technique is suitable for quantification of 5-hmC content in different mouse tissues.