[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-1-en-105":3,"doc-detail-187458-en":53,"doc-seo-187458-105":76},{"code":4,"msg":5,"data":6},0,"success",[7,14,19,24,29,34,39,44,49],{"id":8,"doc_module":9,"doc_module_name":10,"category_name":11,"show_sort_weight":12,"slug":13},11,1,"Template","Presentations",90,"presentations",{"id":15,"doc_module":9,"doc_module_name":10,"category_name":16,"show_sort_weight":17,"slug":18},12,"Resumes",80,"resumes",{"id":20,"doc_module":9,"doc_module_name":10,"category_name":21,"show_sort_weight":22,"slug":23},14,"Invoices",70,"invoices",{"id":25,"doc_module":9,"doc_module_name":10,"category_name":26,"show_sort_weight":27,"slug":28},15,"Posters",60,"posters",{"id":30,"doc_module":9,"doc_module_name":10,"category_name":31,"show_sort_weight":32,"slug":33},16,"Social Media",50,"social-media",{"id":35,"doc_module":9,"doc_module_name":10,"category_name":36,"show_sort_weight":37,"slug":38},17,"Forms",40,"forms",{"id":40,"doc_module":9,"doc_module_name":10,"category_name":41,"show_sort_weight":42,"slug":43},18,"Letters",30,"letters",{"id":45,"doc_module":9,"doc_module_name":10,"category_name":46,"show_sort_weight":47,"slug":48},21,"Paper Templates",5,"papers-templates",{"id":50,"doc_module":9,"doc_module_name":10,"category_name":51,"show_sort_weight":4,"slug":52},158,"General","general-158",{"code":4,"msg":5,"data":54},{"doc_id":55,"user_id":56,"nickname":57,"user_avatar":58,"doc_module":9,"category_id":50,"category_name":51,"doc_title":59,"doc_description":60,"doc_content":61,"file_id":62,"file_url":63,"file_type":64,"file_size":65,"view_count":66,"is_deleted":4,"is_public":9,"is_downloadable":9,"audit_status":9,"page_count":67,"language":68,"language_code":69,"site_id":70,"html_lang":69,"table_of_contents":71,"faqs":72,"seo_title":73,"seo_description":60,"update_tm":74,"read_time":75},187458,2336474466712,"Quinn Holloway","https://ap-avatar.wpscdn.com/davatar_a8503ba1806abce46bf441b54a3ca4cd","dCas9-based transcriptional repression and activation","This study investigates the application of dCas9 (dead Cas9) systems for precise transcriptional regulation in biological systems. The research details the design and implementation of dCas9 fused with two distinct effector domains: RNase III for transcriptional repression and VP64 for activation. The dCas9-RNase III fusion protein, guided by a custom single guide RNA (sgRNA) containing a crRNA and tracrRNA, targets specific DNA sequences to inhibit transcription. This inhibition can occur at the initiation phase or during elongation, as illustrated in Figure 1A. Conversely, the dCas9-VP64 fusion, utilized in conjunction with a transcriptional activator, is employed to enhance gene expression. Experimental data presented in Figures 1B and 1C demonstrate the efficacy of these dCas9-based regulatory systems. Figure 1B shows relative fluorescence measurements indicating a dose-dependent repression or activation effect at different genomic loci. Figure 1C provides Northern blot analysis, confirming the alteration of RNA levels upon dCas9-mediated transcriptional modulation. Further investigations in Figure 2 explore the impact of sgRNA design on repression efficiency, with variations in spacer length and binding sites affecting dCas9 localization and activity. Figure 3 extends this research to a different reporter system (lacZ) and also explores the integration of the omega subunit of RNA polymerase with dCas9 to modulate transcription initiation. The findings collectively highlight the versatility and potential of dCas9 as a powerful tool for programmable gene regulation, enabling precise control over gene expression for various research and biotechnological applications.","","cbCaig2xP2SbBDG3","https://ap.wps.com/l/cbCaig2xP2SbBDG3","pdf",3192330,3,10,"English","en",105,"# dCas9-based transcriptional repression and activation\n## Figure 1: Mechanism and characterization of dCas9-based transcriptional repression and activation\n## Figure 2: Optimization of sgRNA for dCas9-mediated repression\n## Figure 3: dCas9-omega fusion protein for transcriptional modulation","[{\"question\":\"How does the dCas9-RNase III system inhibit transcription?\",\"answer\":\"The dCas9-RNase III system inhibits transcription by binding to a target DNA sequence, guided by an sgRNA, and then degrading the nascent RNA transcript or interfering with the transcriptional machinery, thereby preventing gene expression.\"},{\"question\":\"What is the role of the sgRNA in dCas9-mediated gene regulation?\",\"answer\":\"The sgRNA is crucial for guiding the dCas9 protein to the specific target DNA sequence in the genome. It comprises two parts: the crRNA, which determines the target sequence, and the tracrRNA, which binds to dCas9.\"},{\"question\":\"What is the purpose of fusing the omega subunit to dCas9 in the presented research?\",\"answer\":\"The fusion of the omega subunit of RNA polymerase to dCas9 aims to further enhance or modulate transcriptional regulation by potentially influencing the interaction with the core RNA polymerase complex and affecting transcription initiation efficiency.\"}]","dCas9-based transcriptional repression and activation | PDF",1788383132,4,{"code":4,"msg":77,"data":78},"ok",{"site_id":70,"language":69,"slug":79,"title":59,"keywords":61,"description":60,"schema_data":80,"social_meta":134,"head_meta":136,"extra_data":138,"updated_unix":139},"dcas9-based-transcriptional-repression-and-activation",{"@graph":81,"@context":133},[82,96,116],{"@type":83,"itemListElement":84},"BreadcrumbList",[85,89,92,94],{"item":86,"name":87,"@type":88,"position":9},"https://docshare.wps.com","Home","ListItem",{"item":90,"name":10,"@type":88,"position":91},"https://docshare.wps.com/template/",2,{"item":93,"name":51,"@type":88,"position":66},"https://docshare.wps.com/template/general/",{"item":95,"name":59,"@type":88,"position":75},"https://docshare.wps.com/template/dcas9-based-transcriptional-repression-and-activation/187458/",{"url":95,"name":59,"@type":97,"image":98,"author":103,"headline":59,"publisher":105,"fileFormat":108,"inLanguage":69,"description":60,"dateModified":109,"datePublished":110,"encodingFormat":108,"isAccessibleForFree":111,"interactionStatistic":112},"DigitalDocument",{"url":99,"@type":100,"width":101,"height":102},"https://docshare.wps.com/thumbnails/dcas9-based-transcriptional-repression-and-activation/187458.png","ImageObject",442,249,{"name":57,"@type":104},"Person",{"url":86,"name":106,"@type":107},"DocShare","Organization","application/pdf","2026-09-26","2026-09-02",true,{"@type":113,"interactionType":114,"userInteractionCount":66},"InteractionCounter",{"@type":115},"ViewAction",{"@type":117,"mainEntity":118},"FAQPage",[119,125,129],{"name":120,"@type":121,"acceptedAnswer":122},"How does the dCas9-RNase III system inhibit transcription?","Question",{"text":123,"@type":124},"The dCas9-RNase III system inhibits transcription by binding to a target DNA sequence, guided by an sgRNA, and then degrading the nascent RNA transcript or interfering with the transcriptional machinery, thereby preventing gene expression.","Answer",{"name":126,"@type":121,"acceptedAnswer":127},"What is the role of the sgRNA in dCas9-mediated gene regulation?",{"text":128,"@type":124},"The sgRNA is crucial for guiding the dCas9 protein to the specific target DNA sequence in the genome. It comprises two parts: the crRNA, which determines the target sequence, and the tracrRNA, which binds to dCas9.",{"name":130,"@type":121,"acceptedAnswer":131},"What is the purpose of fusing the omega subunit to dCas9 in the presented research?",{"text":132,"@type":124},"The fusion of the omega subunit of RNA polymerase to dCas9 aims to further enhance or modulate transcriptional regulation by potentially influencing the interaction with the core RNA polymerase complex and affecting transcription initiation efficiency.","https://schema.org",{"og:url":95,"og:type":135,"og:title":59,"og:site_name":106,"og:description":60},"article",{"robots":137,"canonical":95},"index,follow",{"doc_id":55,"site_id":70},1790379272]