{"id":913,"date":"2017-11-13T19:23:17","date_gmt":"2017-11-13T10:23:17","guid":{"rendered":"https:\/\/wp.bs-lab.ks.service.cloud.teu.ac.jp\/?p=913"},"modified":"2018-02-22T15:06:31","modified_gmt":"2018-02-22T06:06:31","slug":"development-of-a-global-dna-methylation-bio-sensing-system-for-cancer-diagnostics","status":"publish","type":"post","link":"https:\/\/yoshida-lab.bs.teu.ac.jp\/?p=913&lang=en","title":{"rendered":"Development of a global DNA methylation bio-sensing system for cancer diagnostics"},"content":{"rendered":"

We developed a bio-sensing system for global DNA methylation levels that can be applied to cancer diagnostics. The paper has been published in Analytica Chimica Acta (Vol. 990, p. 168-173, 2017)1<\/span>.<\/p>\n

DNA methylation of cytosine in CpG dinucleotides has important biological roles in regulation of gene expression. In cancer cells, hypomethylation has been reported in transposable elements, which constitute approximately 45% of the human genome; therefore, global DNA methylation levels are potential biomarkers for cancer diagnostics. We previously developed a bio-sensing system for detecting methylated CpG content in genomic DNA based on bioluminescence resonance energy transfer (BRET) using methyl-CpG-binding domain (MBD)-fused luciferase (MBD-luciferase)2<\/span>. In the system, BRET between the artificial protein and DNA intercalating dye is detected on the methylated CpG site on genomic DNA; thus, methylated CpG content can be detected by the BRET signal (Fig. 1 left). However, a calibration curve was required to quantify global DNA methylation levels since the system does not detect unmethylated CpG content.<\/p>\n

\"\"<\/p>\n

Fig. 1 A bio-sensing system for global DNA methylation levels using CXXC- and MBD-fused luciferase.<\/strong><\/p>\n

We then constructed a new unmethyl-CpG-binding domain (MLL CXXC)-fused luciferase (CXXC-luciferase) and developed a bio-sensing system for detecting unmethylated CpG content on genomic DNA on the same platform (Fig. 1 right). We demonstrated that global DNA methylation levels can be quantified by measuring BRET signals from CXXC-luciferase and MBD-luciferase without a calibration curve. In the future, the bio-sensing system could be applied for home cancer tests.<\/p>\n

1. Yoshida W., Baba Y., Banzawa K., Karube I. A quantitative homogeneous assay for global DNA methylation levels using CpG-binding domain- and methyl-CpG-binding domain-fused luciferase. Anal. Chim. Acta (2017) 990, 168-173
\nhttp:\/\/www.sciencedirect.com\/science\/article\/pii\/S0003267017308504?via%3Dihub<\/a><\/strong><\/p>\n

2. Yoshida W., Baba Y., Karube I. Global DNA Methylation Detection System Using MBD-Fused Luciferase Based on Bioluminescence Resonance Energy Transfer Assay. Anal. Chem. (2016) 88, 9264-9268
\nhttp:\/\/pubs.acs.org\/doi\/abs\/10.1021\/acs.analchem.6b02565<\/a><\/strong><\/p>\n","protected":false},"excerpt":{"rendered":"

We developed a bio-sensing system for global DNA methylation levels that can be applied to cancer diagnostics. […]<\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"sns_share_botton_hide":"","vkExUnit_sns_title":"","_vk_print_noindex":"","sitemap_hide":"","vkExUnit_EyeCatch_disable":"","_veu_custom_css":"","veu_display_promotion_alert":"","vkexunit_cta_each_option":"","twitterCardType":"summary","cardImageID":0,"cardImage":"","cardTitle":"","cardDesc":"","cardImageAlt":"","cardPlayer":"","cardPlayerWidth":0,"cardPlayerHeight":0,"cardPlayerStream":"","cardPlayerCodec":"","_lightning_design_setting":[],"footnotes":"","vk-ltc-link":"","vk-ltc-target":""},"categories":[1],"tags":[],"class_list":["post-913","post","type-post","status-publish","format-standard","hentry","category-uncategorized"],"veu_head_title_object":{"title":"","add_site_title":""},"_links":{"self":[{"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/posts\/913","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=913"}],"version-history":[{"count":4,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/posts\/913\/revisions"}],"predecessor-version":[{"id":1060,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=\/wp\/v2\/posts\/913\/revisions\/1060"}],"wp:attachment":[{"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=913"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=913"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/yoshida-lab.bs.teu.ac.jp\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=913"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}