{"id":477,"date":"2020-01-17T13:59:20","date_gmt":"2020-01-17T13:59:20","guid":{"rendered":"https:\/\/corakid.com\/?page_id=477"},"modified":"2020-04-16T12:26:34","modified_gmt":"2020-04-16T12:26:34","slug":"crisprcas9","status":"publish","type":"page","link":"https:\/\/corakid.com\/en\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/","title":{"rendered":"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9."},"content":{"rendered":"<h4 class=\"wp-block-heading\"><strong>Pr\u00e9sentation<\/strong><\/h4>\n\n\n\n<p>La plateforme utilise la technologie d&rsquo;\u00e9dition du g\u00e9nome \u00e0 l&rsquo;aide du syst\u00e8me CRISPR\/Cas dans le but de g\u00e9n\u00e9rer \u00e0 la demande de la communaut\u00e9 scientifique des clones cellulaires invalid\u00e9s pour des g\u00e8nes d&rsquo;int\u00e9r\u00eat, et des clones cellulaires inductibles pour l&rsquo;expression d&rsquo;un transg\u00e8ne. Selon chaque projet, la plateforme propose une strat\u00e9gie de vectorisation du syst\u00e8me CRISPR-Cas9 ou une transformation par lentivirus, la technique d\u2019\u00e9valuation de l\u2019efficacit\u00e9 et fournit une validation rigoureuse des lign\u00e9es cellulaires fournies. <\/p>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"570\" src=\"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg\" alt=\"http:\/\/www.itun.nantes.inserm.fr\/var\/site\/storage\/images\/_aliases\/full\/2\/0\/7\/3\/3702-2-eng-GB\/GenoCellEdit%2001.png\" class=\"wp-image-479\" srcset=\"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg 1024w, https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-300x167.jpg 300w, https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-768x427.jpg 768w, https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr.jpg 1062w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p>Les\nnucl\u00e9ases artificielles CRISPR\/Cas9 reconnaissent des s\u00e9quences sp\u00e9cifiques\nr\u00e9parties dans tout le g\u00e9nome et g\u00e9n\u00e8rent des cassures de l\u2019ADN. La r\u00e9paration\nde l&rsquo;ADN par NHEJ (non-homologous end joining) est propice aux erreurs qui\ng\u00e9n\u00e8rent des d\u00e9l\u00e9tions et\/ou des insertions de nucl\u00e9otides pouvant produire des\nd\u00e9calages du cadre de lecture provocant ainsi une invalidation de g\u00e8ne\n(knock-out ou KO). La recombinaison homologue (HDR) permet d&rsquo;introduire de\nnouvelles s\u00e9quences dans le g\u00e8ne cibl\u00e9 (knock-in ou KI). Ainsi, les nucl\u00e9ases\nCRISPR\/Cas9 permettent de g\u00e9n\u00e9rer \u00e0 la fois des KO ou des KI.<\/p>\n\n\n\n<p>Les lentivirus\nsont capables d&rsquo;infecter toutes les cellules eukaryotes, y compris les cellules\nen pause r\u00e9plicative. Ces propri\u00e9t\u00e9s et la cr\u00e9ation de syst\u00e8mes lentiviraux\nd\u00e9ficients ne pouvant plus se r\u00e9pliquer procure un moyen efficace simple et s\u00fbr\nde cr\u00e9ation de lign\u00e9es cellulaires transform\u00e9es avec un transg\u00e8ne pouvant \u00eatre\nde grande taille (&gt;10kb). L&rsquo;utilisation du syst\u00e8me Gateway \u00e0 double\ns\u00e9lection (positive + n\u00e9gative) permet de cloner l&rsquo;ADN compl\u00e9mentaire d&rsquo;un g\u00e8ne\ndans un vecteur d&rsquo;expression avec une efficacit\u00e9 proche de 100%.<\/p>\n\n\n\n<h4 class=\"wp-block-heading\"><strong>Expertise<\/strong><\/h4>\n\n\n\n<p>Notre expertise porte sur la strat\u00e9gie de\nciblage adapt\u00e9e \u00e0 chaque g\u00e8ne, le design des sgRNAs, l&rsquo;appr\u00e9ciation du off\ntarget, sur la transfection de lign\u00e9es cellulaires eukaryotes ainsi que sur la\nvalidation des clones cellulaires produits (validation de la mutation pour le\nCrispr\/Cas9, et validation d&rsquo;un clone inductible avec expression basale\nminimale du transg\u00e8ne pour les clones inductibles transform\u00e9s par lentivirus). <\/p>\n\n\n\n<h4 class=\"wp-block-heading\"><strong>Service et engagement<\/strong><\/h4>\n\n\n\n<p>\u00c9tudier vos besoins et effectuer l\u2019analyse de faisabilit\u00e9 et le\nd\u00e9veloppement m\u00e9thodologique.<\/p>\n\n\n\n<p>Produire des clones cellulaires invalid\u00e9s et\/ou inductibles par la m\u00e9thode\nCRISPR\/Cas9 ou par transformation lentivirale.<\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Ouvert aux \u00e9tablissements publics (prioritaires) et priv\u00e9s<\/li><\/ul>\n\n\n\n<h4 class=\"wp-block-heading\"><strong>Services<\/strong><\/h4>\n\n\n\n<h5 class=\"wp-block-heading\">Crispr<\/h5>\n\n\n\n<ul class=\"wp-block-list\"><li>Etude du g\u00e8ne cible et des strat\u00e9gies possibles<\/li><li>Strat\u00e9gie, le type de Cas, les vecteurs et leurs utilisations<\/li><li>Design <em>in silico<\/em> des sgRNA ciblant le site g\u00e9nomique d\u2019int\u00e9r\u00eat<\/li><li>Appr\u00e9ciation du off target<\/li><li>Vectorisation de l\u2019outil (plasmides, vecteurs lentiviraux)<\/li><li>Validation fonctionnelle des sgRNA<\/li><li>Design et construction du donneur ADN pour modifier \u00e0 fa\u00e7on le g\u00e9nome et sa d\u00e9tection (pour les knock in)<\/li><li>Mise au point de nouvelles techniques d&rsquo;\u00e9dition de g\u00e8ne (Cas12, etc.)<\/li><\/ul>\n\n\n\n<h5 class=\"wp-block-heading\">Lign\u00e9es inductibles<\/h5>\n\n\n\n<ul class=\"wp-block-list\"><li>Etude du g\u00e8ne cible et choix de l&rsquo;isoforme \u00e0 cloner<\/li><li>clonage du g\u00e8ne cible dans un vecteur d&rsquo;expression\ninductible<\/li><li>production de lentivirus<\/li><li>transformation lentivirale<\/li><li>isolation de clones cellulaires<\/li><li>caract\u00e9risation fonctionnelle de clones cellulaire\n(expression basale et induction du transg\u00e8ne)<\/li><\/ul>","protected":false},"excerpt":{"rendered":"<p>Pr\u00e9sentation La plateforme utilise la technologie d&rsquo;\u00e9dition du g\u00e9nome \u00e0 l&rsquo;aide du syst\u00e8me CRISPR\/Cas dans le but de g\u00e9n\u00e9rer \u00e0 la demande de la communaut\u00e9 scientifique des clones cellulaires invalid\u00e9s pour des g\u00e8nes d&rsquo;int\u00e9r\u00eat, et des clones cellulaires inductibles pour l&rsquo;expression d&rsquo;un transg\u00e8ne. Selon chaque projet, la plateforme propose une strat\u00e9gie de vectorisation du syst\u00e8me [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":233,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_cbd_carousel_blocks":"[]","footnotes":""},"class_list":["post-477","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.5 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9. - CoRaKiD<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/corakid.com\/en\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9. - CoRaKiD\" \/>\n<meta property=\"og:description\" content=\"Pr\u00e9sentation La plateforme utilise la technologie d&rsquo;\u00e9dition du g\u00e9nome \u00e0 l&rsquo;aide du syst\u00e8me CRISPR\/Cas dans le but de g\u00e9n\u00e9rer \u00e0 la demande de la communaut\u00e9 scientifique des clones cellulaires invalid\u00e9s pour des g\u00e8nes d&rsquo;int\u00e9r\u00eat, et des clones cellulaires inductibles pour l&rsquo;expression d&rsquo;un transg\u00e8ne. Selon chaque projet, la plateforme propose une strat\u00e9gie de vectorisation du syst\u00e8me [&hellip;]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/corakid.com\/en\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/\" \/>\n<meta property=\"og:site_name\" content=\"CoRaKiD\" \/>\n<meta property=\"article:modified_time\" content=\"2020-04-16T12:26:34+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Estimated reading time\" \/>\n\t<meta name=\"twitter:data1\" content=\"3 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/\",\"url\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/\",\"name\":\"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\\\/Cas9. - CoRaKiD\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#website\"},\"primaryImageOfPage\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/#primaryimage\"},\"image\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/#primaryimage\"},\"thumbnailUrl\":\"https:\\\/\\\/corakid.com\\\/wp-content\\\/uploads\\\/2020\\\/01\\\/image-crispr-1024x570.jpg\",\"datePublished\":\"2020-01-17T13:59:20+00:00\",\"dateModified\":\"2020-04-16T12:26:34+00:00\",\"breadcrumb\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/#breadcrumb\"},\"inLanguage\":\"en-GB\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/\"]}]},{\"@type\":\"ImageObject\",\"inLanguage\":\"en-GB\",\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/#primaryimage\",\"url\":\"https:\\\/\\\/corakid.com\\\/wp-content\\\/uploads\\\/2020\\\/01\\\/image-crispr-1024x570.jpg\",\"contentUrl\":\"https:\\\/\\\/corakid.com\\\/wp-content\\\/uploads\\\/2020\\\/01\\\/image-crispr-1024x570.jpg\"},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/crisprcas9\\\/#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Accueil\",\"item\":\"https:\\\/\\\/corakid.com\\\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"Plateformes\",\"item\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/\"},{\"@type\":\"ListItem\",\"position\":3,\"name\":\"G\u00e9nomique et Biologie cellulaire\",\"item\":\"https:\\\/\\\/corakid.com\\\/core-facilities\\\/genomique-et-biologie-cellulaire\\\/\"},{\"@type\":\"ListItem\",\"position\":4,\"name\":\"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\\\/Cas9.\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#website\",\"url\":\"https:\\\/\\\/corakid.com\\\/en\\\/\",\"name\":\"CoRaKiD\",\"description\":\"UMRS 1155 Common and Rare Kidney Diseases\",\"publisher\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#organization\"},\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\\\/\\\/corakid.com\\\/en\\\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"en-GB\"},{\"@type\":\"Organization\",\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#organization\",\"name\":\"CoRaKiD\",\"url\":\"https:\\\/\\\/corakid.com\\\/en\\\/\",\"logo\":{\"@type\":\"ImageObject\",\"inLanguage\":\"en-GB\",\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#\\\/schema\\\/logo\\\/image\\\/\",\"url\":\"https:\\\/\\\/corakid.com\\\/wp-content\\\/uploads\\\/2019\\\/10\\\/logo5.png\",\"contentUrl\":\"https:\\\/\\\/corakid.com\\\/wp-content\\\/uploads\\\/2019\\\/10\\\/logo5.png\",\"width\":652,\"height\":247,\"caption\":\"CoRaKiD\"},\"image\":{\"@id\":\"https:\\\/\\\/corakid.com\\\/en\\\/#\\\/schema\\\/logo\\\/image\\\/\"}}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9. - CoRaKiD","robots":{"index":"index","follow":"follow","max-snippet":"max-snippet:-1","max-image-preview":"max-image-preview:large","max-video-preview":"max-video-preview:-1"},"canonical":"https:\/\/corakid.com\/en\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/","og_locale":"en_GB","og_type":"article","og_title":"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9. - CoRaKiD","og_description":"Pr\u00e9sentation La plateforme utilise la technologie d&rsquo;\u00e9dition du g\u00e9nome \u00e0 l&rsquo;aide du syst\u00e8me CRISPR\/Cas dans le but de g\u00e9n\u00e9rer \u00e0 la demande de la communaut\u00e9 scientifique des clones cellulaires invalid\u00e9s pour des g\u00e8nes d&rsquo;int\u00e9r\u00eat, et des clones cellulaires inductibles pour l&rsquo;expression d&rsquo;un transg\u00e8ne. Selon chaque projet, la plateforme propose une strat\u00e9gie de vectorisation du syst\u00e8me [&hellip;]","og_url":"https:\/\/corakid.com\/en\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/","og_site_name":"CoRaKiD","article_modified_time":"2020-04-16T12:26:34+00:00","og_image":[{"url":"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg","type":"","width":"","height":""}],"twitter_card":"summary_large_image","twitter_misc":{"Estimated reading time":"3 minutes"},"schema":{"@context":"https:\/\/schema.org","@graph":[{"@type":"WebPage","@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/","url":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/","name":"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9. - CoRaKiD","isPartOf":{"@id":"https:\/\/corakid.com\/en\/#website"},"primaryImageOfPage":{"@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/#primaryimage"},"image":{"@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/#primaryimage"},"thumbnailUrl":"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg","datePublished":"2020-01-17T13:59:20+00:00","dateModified":"2020-04-16T12:26:34+00:00","breadcrumb":{"@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/#breadcrumb"},"inLanguage":"en-GB","potentialAction":[{"@type":"ReadAction","target":["https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/"]}]},{"@type":"ImageObject","inLanguage":"en-GB","@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/#primaryimage","url":"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg","contentUrl":"https:\/\/corakid.com\/wp-content\/uploads\/2020\/01\/image-crispr-1024x570.jpg"},{"@type":"BreadcrumbList","@id":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/crisprcas9\/#breadcrumb","itemListElement":[{"@type":"ListItem","position":1,"name":"Accueil","item":"https:\/\/corakid.com\/"},{"@type":"ListItem","position":2,"name":"Plateformes","item":"https:\/\/corakid.com\/core-facilities\/"},{"@type":"ListItem","position":3,"name":"G\u00e9nomique et Biologie cellulaire","item":"https:\/\/corakid.com\/core-facilities\/genomique-et-biologie-cellulaire\/"},{"@type":"ListItem","position":4,"name":"G\u00e9n\u00e9rations de clones cellulaires(toutes esp\u00e8ces) invalid\u00e9s et inductibles par la technologie CRISPR\/Cas9."}]},{"@type":"WebSite","@id":"https:\/\/corakid.com\/en\/#website","url":"https:\/\/corakid.com\/en\/","name":"CoRaKiD","description":"UMRS 1155 Common and Rare Kidney Diseases","publisher":{"@id":"https:\/\/corakid.com\/en\/#organization"},"potentialAction":[{"@type":"SearchAction","target":{"@type":"EntryPoint","urlTemplate":"https:\/\/corakid.com\/en\/?s={search_term_string}"},"query-input":{"@type":"PropertyValueSpecification","valueRequired":true,"valueName":"search_term_string"}}],"inLanguage":"en-GB"},{"@type":"Organization","@id":"https:\/\/corakid.com\/en\/#organization","name":"CoRaKiD","url":"https:\/\/corakid.com\/en\/","logo":{"@type":"ImageObject","inLanguage":"en-GB","@id":"https:\/\/corakid.com\/en\/#\/schema\/logo\/image\/","url":"https:\/\/corakid.com\/wp-content\/uploads\/2019\/10\/logo5.png","contentUrl":"https:\/\/corakid.com\/wp-content\/uploads\/2019\/10\/logo5.png","width":652,"height":247,"caption":"CoRaKiD"},"image":{"@id":"https:\/\/corakid.com\/en\/#\/schema\/logo\/image\/"}}]}},"_links":{"self":[{"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/pages\/477","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/comments?post=477"}],"version-history":[{"count":2,"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/pages\/477\/revisions"}],"predecessor-version":[{"id":620,"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/pages\/477\/revisions\/620"}],"up":[{"embeddable":true,"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/pages\/233"}],"wp:attachment":[{"href":"https:\/\/corakid.com\/en\/wp-json\/wp\/v2\/media?parent=477"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}