{"id":1065,"date":"2024-09-13T15:44:25","date_gmt":"2024-09-13T15:44:25","guid":{"rendered":"http:\/\/www.caltagmedsystems.co.uk\/information\/?p=1065"},"modified":"2025-06-13T11:10:09","modified_gmt":"2025-06-13T11:10:09","slug":"fluorescent-ubiquitination-based-cell-cycle-indicator-fucci","status":"publish","type":"post","link":"https:\/\/www.caltagmedsystems.co.uk\/information\/fluorescent-ubiquitination-based-cell-cycle-indicator-fucci\/","title":{"rendered":"FUCCI: Fluorescent Ubiquitination-based Cell Cycle Indicator"},"content":{"rendered":"\n<p>FUCCI (Fluorescent Ubiquitination-based Cell Cycle Indicator) is a set of fluorescent probes which enables the visualisation of cell cycle progression in living cells. FUCCI utilises the phase-dependent nature of replication licensing factors Cdt1 and Geminin. A fusion protein of a fragment of Cdt1 (amino acids 30-120) with the fluorescent protein monomeric Kusabira-Orange 2 (mKO2) serves as an indicator of the G1 phase. A fusion protein of a fragment of Geminin (amino acids 1-110 or 1-60) with the fluorescent protein monomeric Azami-Green 1 (mAG1) visualises the S, G2 and M phases. In summary, FUCCI utilises the highly selective, rapid degradation of the replication licensing factors mediated by the ubiquitin proteasome system to give excellent visualisations of the cell cycle.<\/p>\n\n\n\n<p><strong>Cdt1: <\/strong>Cdc10 dependent transcript 1 is a conserved replication factor required for licensing the chromosome for a single course of DNA synthesis. Abundantly expressed throughout the cell cycle, Cdt1 is ubiquitinated by the ubiqutin ligase complex SCFskp2 during S and G2 phases and degraded by proteasome.<\/p>\n\n\n\n<p><strong>Geminin: <\/strong>Geminin inhibits the licensing activity of Cdt1. Geminin interferes with the binding of licensing factors to the replication origin once a chromosome has started to replicate during the S phase. During M and G1 phases, Geminin is ubiquitinated by the E3 ligase complex APCcdh1 and degraded by the proteasome.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">What makes FUCCI a powerful research tool?<\/h2>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Real-time visualisation of cell cycle progression<\/li>\n\n\n\n<li>Spatio-temporal imaging of cell cycle dynamics<\/li>\n\n\n\n<li>Utilisation of fluorescent proteins Green mAG1 and Orange mKO2<\/li>\n\n\n\n<li>Fucci-G1 Orange labels G1 phase nuclei in orange. Fucci-S\/G2\/M Green labels S\/G2\/M phases nuclei in green.<\/li>\n<\/ul>\n\n\n\n<p>By visualising the cell cycle, Fucci is a powerful tool to investigate any process that has to do with cell growth and differentiation, such as the development and regeneration of organs as well as carcinogenesis.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Fluorescent Images of Fucci Cells<\/h3>\n\n\n\n<figure class=\"wp-block-image size-full\"><img loading=\"lazy\" decoding=\"async\" width=\"242\" height=\"212\" src=\"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-content\/uploads\/image-301.png\" alt=\"\" class=\"wp-image-10970\"\/><\/figure>\n\n\n\n<p>Each cell cycle of the G1, G1\/S, S, G2, and M phases can be determined by the combination of Fucci-G1 Orange, Fucci-S\/G2\/M Green, and an antibody against PCNA (Proliferation Cell Nuclear Antigen). The G1 phase is indicated by orange. Both orange and green were observed in the G1\/S phase. Additional immunostaining colour by PCNA was observed at the initiation of the S phase. Cells with pure green fluorescence were either in the S or G2 phase and were distinguished by immunostaining of the S phase. The rest of the cells were classified into the M phase. These results are consistent with the fact that Cdt1 accumulates in G1, while Geminin accumulates in S\/G2\/M. (Provided by Dr. Sakaue-Sawano at RIKEN. Cell (2008) 132:487-98).<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Applications<\/h2>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>High Content Imaging<\/strong> &#8211; FUCCI can be used for the visualisation and analysis of single-cell metrics which can provide rich data for analysis.<\/li>\n\n\n\n<li><strong>Cancer Inhibitor Screening<\/strong> &#8211; FUCCI was used to test the effect of selinexor on cancer cell growth. After imaging the cell cycle effects of selinexor using FUCCI, it was determined that G-1 or early S-phase treated cells show the strongest response and most often die or arrest.<\/li>\n<\/ul>\n\n\n\n<div class=\"wp-block-buttons is-content-justification-center is-layout-flex wp-container-core-buttons-is-layout-a89b3969 wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button\"><a class=\"wp-block-button__link has-text-color has-background has-link-color wp-element-button\" href=\"https:\/\/www.caltagmedsystems.co.uk\/pricing_ordering\/?group_1=All&amp;group_2=All&amp;supplier=10&amp;clonality=All&amp;host=All&amp;species=All&amp;applications=All&amp;regulatory_status=All&amp;searchwords=fucci\" style=\"color:#fbc100;background-color:#21318f\"><strong>See all FUCCI products<\/strong><\/a><\/div>\n<\/div>\n\n\n\n<p class=\"clear\"><\/p>\n\n\n\n<hr class=\"wp-block-separator has-text-color has-alpha-channel-opacity has-background\" style=\"background-color:#21318f;color:#21318f\"\/>\n\n\n\n<p>Originally posted on:\u00a0<a href=\"https:\/\/www.mblbio.com\/bio\/g\/product\/flprotein\/fucci.html\">https:\/\/www.mblbio.com\/bio\/g\/product\/flprotein\/fucci.html<\/a><\/p>\n\n\n\n<p><a href=\"https:\/\/www.caltagmedsystems.co.uk\/\">Caltag Medsystems&nbsp;<\/a>is the distributor of&nbsp;<a href=\"https:\/\/www.caltagmedsystems.co.uk\/mbl\/\">MBL&nbsp;<\/a>products in the UK and Ireland. If you have any questions about these products, please&nbsp;<a href=\"https:\/\/www.caltagmedsystems.co.uk\/contact.php\">contact us<\/a>.<\/p>\n\n\n\n<p><\/p>\n","protected":false},"excerpt":{"rendered":"<p>FUCCI (Fluorescent Ubiquitination-based Cell Cycle Indicator) is a set of fluorescent probes which enables the visualisation of cell cycle progression in living cells.<\/p>\n","protected":false},"author":13,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[73,681],"tags":[53,66,939,67],"class_list":["post-1065","post","type-post","status-publish","format-standard","hentry","category-drug-discovery","category-proteins","tag-cell-cycle","tag-drug-discovery","tag-fucci","tag-mbl-international"],"_links":{"self":[{"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/posts\/1065","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/users\/13"}],"replies":[{"embeddable":true,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/comments?post=1065"}],"version-history":[{"count":8,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/posts\/1065\/revisions"}],"predecessor-version":[{"id":12317,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/posts\/1065\/revisions\/12317"}],"wp:attachment":[{"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/media?parent=1065"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/categories?post=1065"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.caltagmedsystems.co.uk\/information\/wp-json\/wp\/v2\/tags?post=1065"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}