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cookie exists means show popup. */ cell_phone_info = '<br><a href="'+my_account+'" class="cell-phone-details">Click here</a> to save Cell Phone Number.'; } var cookie_data = tcpee_readCookie('cc_popup_shown'); /* END: show enter credit card bar */ /* START: hide popup */ jQuery(".close_me_for_now").on("click", function(e){ jQuery(".save_cc_details_container").hide(); jQuery("#light_box_fade_div").hide(); }); jQuery("#light_box_fade_div").on("click", function(e){ jQuery(".save_cc_details_container").hide(); jQuery("#light_box_fade_div").hide(); }); /* hide and create cookie. */ jQuery(".dont_bug_me").on("click", function(e){ jQuery(".save_cc_details_container").hide(); jQuery("#light_box_fade_div").hide(); tcpee_createCookie("cc_popup_shown", "yes", 0); }); /* END: hide popup */ /* remove anchor notifiaction from single auction product page */ /* jQuery('.single-product .woocommerce-error li a.button, a.save-phone-details').removeAttr('href'); */ /* show login popup on auction single 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Please log into your account to place a bid.</h4><form class="woocomerce-form woocommerce-form-login login" method="post"><p class="woocommerce-form-row woocommerce-form-row--wide form-row form-row-wide"><label for="username">Username <span class="required">*</span></label><input type="text" class="woocommerce-Input woocommerce-Input--text input-text" name="username" id="username" value="" /></p><p class="woocommerce-form-row woocommerce-form-row--wide form-row form-row-wide"><label for="password">Password <span class="required">*</span></label><input class="woocommerce-Input woocommerce-Input--text input-text" type="password" name="password" id="password" /></p><p class="woocommerce-Button-login form-row"><input type="hidden" id="woocommerce-login-nonce" name="woocommerce-login-nonce" value="9454b18658" /><input type="hidden" name="_wp_http_referer" value="/research-resources/" /><input type="submit" class="woocommerce-Button button" name="login" value="Login" /></p><p class="mb-0 woocommerce-LostPassword lost_password text-center"><a href="https://www.emilysentourage.org/?page_id=8118">Register User</a></p><p class="woocommerce-LostPassword lost_password text-center"><a href="https://www.emilysentourage.org/emilyadmin/?action=lostpassword">Forgot your password?</a></p></form><div class="tickets-page-sec"><div class="bottom-style-box p-3"><h3 class="mb-2">Having Technical Difficulties?</h3><p class="mb-0">Please contact us at <a class="tickets-page-sec-link" href="mailto:events@emilysentourage.org">events@emilysentourage.org</a> or call Nick Dinges at <a class="tickets-page-sec-link" href="tel:610-324-1341">610-324-1341</a> for assistance.</p></div></div></div><span class="close_me_for_now" id="close_this_light_box">X</span></div></div><div id="light_box_fade_div" class="common_class_to_close"></div>'); } }); jQuery(document).on("click", ".text-alerts-popup .close_me_for_now, #light_box_fade_div", function(e){ jQuery('.text-alerts-popup, #light_box_fade_div').hide(); }); /* show cell conormation on dashborad and auction single page*/ jQuery('.text-phone-alert, a.save-phone-details').click(function(e) { e.preventDefault(); jQuery('body').prepend('<div class="text-alerts-popup cell-phone-popup save_cc_details_container"><div class="update_phone_number_container update-otp-phone"><h4>Uh oh! Looks like you aren’t signed up to get auction text alerts.</h4><div class="phone-noti-icon"><img src="https://www.emilysentourage.org/wp-content/plugins/tickera-custom-plugin/assets/img/phone-notification-icon.png"></div><h4>Please add your cell phone number.</h4><div class="phone-box-input">+1 <input type="phone" id="update_phone_txt" class="update_phone_txt text-field" value="" maxlength="14" placeholder="Phone number" /><span class="button-loader"><input type="button" id="update_phone_btn" class="update_phone_btn text-field-update" value="Submit" /><img src="https://www.emilysentourage.org/wp-content/plugins/tickera-custom-plugin/assets/img/ajax_loader.gif" class="update_loader_img ajax_loader" style="display:none;" /></span></div></div><div class="update_otp_container update-otp-phone"><h4 class="mt-5">You will receive a text message with a verification code. 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id="close_this_light_box">X</span></div><div id="light_box_fade_div" class="common_class_to_close"></div>'); }); var total_bid_item = jQuery(".bid-box .bid-box-text-move .bid-box-item").size(); var item_height = jQuery(".bid-box .bid-box-text-move .bid-box-item").outerHeight(); var scroll_container_height = ( ( item_height )* total_bid_item ); jQuery(".bid-box .bid-box-text-move").css("height", scroll_container_height); var item_index = 1; var timer = setInterval( function(){ jQuery(".bid-box .bid-box-text-move").animate({ "margin-top": (-1) * ( (item_height) * item_index)} , 2500); if (item_index != ( total_bid_item - 1 )) { item_index = item_index + 1 ; } else { item_index = 0; } }, 5000); }); </script> </head> <body class="page-template-default page page-id-13172 theme-charityhub woocommerce-no-js translatepress-en_US elementor-default elementor-kit-21184">   <div class="body-wrapper page_with_title_cls donate_page_wrapper_cls" data-home="https://www.emilysentourage.org" > <header 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class="gdlr-page-title-container container" > <div class="gdlr-page-title-inner" > <h1 class="gdlr-page-title">Research Resources</h1> </div> </div> </div>  </header> <div class="content-wrapper dasdas"> <div class="gdlr-content">  <div class="above-sidebar-wrapper"><section id="content-section-1" ><div class="section-container container"><div class="twelve columns" ><div class="gdlr-item gdlr-content-item" style="margin-bottom: 40px;" ><p><span style="font-weight: 400;">Robust, validated cell, disease and animal models are critical to accelerate research and drug discovery and development for cystic fibrosis (CF)</span><span style="font-weight: 400;">. A lack of model systems </span><span style="font-weight: 400;">to support investigation of CFTR mutations</span><span style="font-weight: 400;"> has previously served as a barrier to progress. At Emily’s Entourage, we are deeply committed to investing in and developing the most rigorously tested model systems and research resources for individuals with CF that do not benefit from current CFTR modulators. </span></p> <p><span style="font-weight: 400;">Resources available to the research community include:</span></p> <h3><strong>Complementary DNA Constructs</strong></h3> <p><span style="font-weight: 400;">cDNA constructs expressing mutated (W1282X-CFTR) and truncated CFTR produced by the W1282X-CFTR mutation (CFTR<sub>1281</sub></span><span style="font-weight: 400;">) are available. Constructs were generated in pcDNA3.1/Zeo(+), and can be readily subcloned into alternative vectors. Further information is available <a href="http://www.jbc.org/content/292/3/771.full?sid=5f4a2350-e537-4c6e-bcba-79447a4cde03" target="_blank" rel="noopener">here</a>. Generation of these cDNA constructs was supported by Emily’s Entourage. </span></p> <hr /> <h3><strong>Viral Constructs</strong></h3> <p>Lentiviral and adenoviral constructs encoding anticodon-engineered transfer-RNAs (ACE-tRNAs) that recognize and efficiently suppress CFTR premature termination codons (PTCs) genotypes, including W1282X, have been generated by John Lueck, PhD, University of Rochester Medical Center. Further information is available <a href="https://www.biorxiv.org/content/10.1101/2024.06.06.597760v1" target="_blank" rel="noopener">here</a>. The generation of these viral constructs was supported by Emily’s Entourage.</p> <hr /> <h3><strong>Cell Models</strong></h3> <p><strong>Airway epithelial cells<br /> </strong></p> <ul> <li>A panel of airway epithelial cells was generated by Finn J. Hawkins, MBBCh, Boston University, using induced pluripotent stem cells (iPSCs) from pwCF with W1282X-CFTR (and additional CFTR mutations). These cells can be maintained in culture, differentiated into pseudostratified epithelium in air liquid interface for electrophysiological assessments, and cryopreserved. This is a valuable research tool for studying CF and accelerating therapeutic development for CF caused by rare variants. The development of this panel of cells was supported by Emily’s Entourage. For specific genotype availability and to initiate MTA requests, don’t hesitate to contact Dr. Finn J. Hawkins.</li> </ul> <p><strong>Engineered HEK293T cells </strong></p> <ul> <li>Human Embryonic Kidney (HEK) 293T cells expressing 3HA-tagged W1282X-CFTR (and additional CFTR mutations) are available from Marianne Carlon, PhD, KU Leuven. These cell lines are a great research tool for rapid screening of many CFTR correction strategies (e.g., compounds, gene addition, and gene editing), are easily transfected, and the 3HA-tag allows straightforward readout on protein level (e.g. by Western Blot and immunocytochemistry for plasma membrane localization). The generation of these engineered cell lines has been published in open-access format (Bulcaen et al., Cell Reports Medicine, 2024) and was supported by Emily’s Entourage. For specific genotype availability and to initiate MTA requests, don’t hesitate to contact Dr. Marianne Carlon.</li> <li>A previously described prime editing reporter (PEAR; Simon et al., eLife, 2022) has been adapted by Marianne Carlon, PhD, KU Leuven, for compatibility with lentiviral particle production. A stable cell line (PEAR cell line) encoding the GFP reporter with disrupted splice site, in combination with constitutive expression of miRFP was generated. Upon successful prime or adenine base editing, the PEAR-HEK293T cell line becomes GFP-positive, allowing the investigation of new PE- or ABE-VLP architectures, optimisation production protocols and functionally titrating new preps. The generation of this stable cell line was supported by Emily’s Entourage. To initiate MTA requests, don’t hesitate to contact Dr. Marianne Carlon.</li> </ul> <p><strong>Rat thyroid epithelial cell models</strong></p> <ul> <li><span style="font-weight: 400;">Fischer rat thyroid (FRT) cell models expressing W1282X-CFTR and CFTR</span><span style="font-weight: 400;"><sub>1281</sub></span><span style="font-weight: 400;"> (with the EYFP-H148Q/I152L/F46L halide-sensitive reporter). These cell models are amenable to fluorescence and electrophysiological assessment of CFTR activity. Further information is available <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5247652/" target="_blank" rel="noopener">here</a>. Generation of these cell models was supported by Emily’s Entourage.</span></li> </ul> <p><strong>Immortalized airway epithelial cell models </strong></p> <ul> <li><span style="font-weight: 400;">Immortalized CFBE41o- airway epithelial cells expressing W1282X-CFTR or CFTR</span><span style="font-weight: 400;"><sub>1281</sub></span><span style="font-weight: 400;"> with engineered triplet HA epitope tags, to facilitate biochemistry and cell surface presentation, have been generated by Gergerly Lukacs, MD, PhD, McGill University. </span></li> </ul> <p><strong>Gene-edited airway epithelial cell models</strong></p> <ul> <li><span style="font-weight: 400;">Gene edited immortalized 16HBE14o- human bronchial epithelial cells expressing W1282X-CFTR (and additional CFTR mutations) have been generated by the Cystic Fibrosis Foundation Therapeutics Lab and is available <a href="https://www.emilysentourage.org/wp-content/uploads/Valley_JCF_2019.pdf" target="_blank" rel="noopener">here</a>.<br /> </span></li> <li>The gene-edited 16HBE14o- cells expressing W1282X-CFTR (and additional CFTR mutations) generated by the CFFT Lab have been engineered by Marianne Carlon, PhD, KU Leuven to express halide-sensitive (HS) yellow fluorescence protein (YFP). The stable expression of HS-YFP in these cells allows functional CFTR correction to be monitored. It can be a valuable research tool to investigate the efficacy of gene editing or other correction strategies. The engineering of these cell models was supported by Emily’s Entourage. For specific genotype availability and to initiate MTA requests, don’t hesitate to contact Dr. Marianne Carlon.</li> </ul> <p><strong>Growth enhanced airway epithelial cell models </strong></p> <ul> <li><span style="font-weight: 400;">Near-native, growth-enhanced homozygous W1282X-CFTR nasal epithelial cells expressing a proto-oncogene (Bmi-1) and the catalytic subunit of telomerase (hTERT) are available from Scott Randell, PhD, University of North Carolina, Chapel Hill. Further information is available <a href="https://www.emilysentourage.org/wp-content/uploads/UNC_CFTRW1282X_cell_model.pdf" target="_blank" rel="noopener">here</a>. Generation of these cell models was supported by Emily’s Entourage. </span></li> </ul> <p><b>Human primary airway epithelial cells and intestinal organoids</b></p> <ul> <li><span style="font-weight: 400;">Primary nasal, lung, and intestinal cells with two nonsense mutations (including homozygous W1282X/W1282X cells) are available from CFF’s RARE cell collection at the Cystic Fibrosis Foundation Therapeutics (CFFT) Lab. Cells will be distributed based on availability and evaluation of a short (~half page) research plan that will be evaluated for scientific merit. For specific genotype availability, please contact Jessica Stach. MTA requests can be initiated via the CFFT Lab </span><a href="http://www.cff.org/LabMaterialRequest" target="_blank" rel="noopener"><span style="font-weight: 400;">here</span></a><span style="font-weight: 400;">.</span></li> </ul> <p><b>Human primary nasal epithelial cells and stem cell models</b></p> <ul> <li>A biobank of nasal epithelial cells and induced pluripotent stem cells (iPSCs) isolated from individuals living with CF, including those homozygous or heterozygous for the W1282X and/or G542X mutations, is available from The CF Canada-SickKids Program in Individual Therapy (<a href="https://lab.research.sickkids.ca/cfit/cystic-fibrosis-patients-families-researchers/cell-resources-available/" target="_blank" rel="noopener">CFIT</a>). Certain iPSCs have been CrispR-edited to generate isogenic controls.</li> </ul> <p><strong>Rectal organoids</strong></p> <ul> <li><span style="font-weight: 400;">Rectal organoids derived from homozygous W1282X-CFTR subjects are maintained by and available from <a href="https://hub4organoids.eu/" target="_blank" rel="noopener">Hubrecht Organoid Technology</a></span><span style="font-weight: 400;"> and the <a href="https://www.cff.org/LabMaterialRequest/" target="_blank" rel="noopener">Cystic Fibrosis Foundation Therapeutics Lab</a>.</span></li> <li>Patient-derived intestinal organoids (PDIOS) stable expressing ACE-tRNAs have been developed via lentiviral transduction by Jeffrey M. Beekman, PhD, UMC Utrecht. Developing these cell lines will benefit specific experiments such as qPCRs, Western-Blots, and toxicity-related assays. Further information is available <a href="https://www.biorxiv.org/content/10.1101/2024.06.06.597760v1" target="_blank" rel="noopener">here</a>. Emily’s Entourage supported the generation of stable PDIOS-expressing ACE-tRNAs.</li> </ul> <p><strong>Stem cell model</strong></p> <ul> <li><span style="font-weight: 400;">Induced pluripotent stem cells homozygous for the W1282X-CFTR mutation are available from the <a href="https://www.seas.upenn.edu/~diamond/CellLines.html" target="_blank" rel="noopener">Diamond Lab, University of Pennsylvania</a>. </span></li> </ul> <hr /> <h3><strong>Machine-learning-based Algorithms</strong></h3> <ul> <li>A machine-learning-based algorithm named DETECTOR for CF organoid analysis has been developed by Marianne Carlon, PhD, KU Leuven and has been described <a href="https://www.cell.com/cell-reports-medicine/fulltext/S2666-3791(24)00234-9?_returnURL=https://linkinghub.elsevier.com/retrieve/pii/S2666379124002349?showall=true" target="_blank" rel="noopener">here</a> (Bulcaen et al., Cell Reports Medicine, 2024). DETECTOR is a tool for rapid testing and analysis of gene editing therapy approaches in intestinal organoids. Despite being developed explicitly for gene editing work in organoids, this tool can easily be applied to gene addition or any other strategy leading to functional CFTR correction. The generation of this organoid tool was supported by Emily’s Entourage and is freely available through <a href="https://github.com/RL-arch/detector" target="_blank" rel="noopener">GitHub.</a></li> </ul> <hr /> <h3><strong>Murine Models</strong></h3> <ul> <li><span style="font-weight: 400;">Murine models to investigate Cystic Fibrosis (CF) pathology, including models of CF caused by nonsense mutations are available from Craig Hodges, PhD, at the <a href="http://www.cfmice.org/" target="_blank" rel="noopener">Case Western Reserve University Cystic Fibrosis Mouse Model Core</a></span><span style="font-weight: 400;">.</span></li> </ul> <hr /> <h3><strong>Nanoparticles</strong></h3> <p><strong>Lipid Nanoparticles</strong></p> <ul> <li>Novel lipid nanoparticles encapsulating RNA (mRNA and sgRNA) have been developed by the laboratory of Debadyuti (Rana) Ghosh, PhD, The University of Texas at Austin. The generation of this formulation was supported by Emily’s Entourage, and its composition and methods for formulation are available for other research groups to deliver RNA therapeutic cargo. To initiate MTA requests, don’t hesitate to contact Dr. Ghosh.<strong><br /> </strong></li> </ul> <hr /> <p>For questions about these resources, please <a href="https://www.emilysentourage.org/research-inquiries/" target="_blank" rel="noopener">Contact Us.</a></p> </div></div><div class="clear"></div></div></section></div>   </div> <div class="clear" ></div> </div> <footer class="footer-wrapper donate_footer" > <div class="footer-container container row text-center"> <div class="footer-column col-md-3" id="footer-widget-1"><div id="custom_html-5" class="widget_text widget widget_custom_html gdlr-item gdlr-widget"><div class="textwidget custom-html-widget"><ul class="footer_end_logo"><li class="guide-star-logo"><a href="https://www.guidestar.org/profile/45-3768161" target="_blank" rel="noopener"><img src="https://widgets.guidestar.org/gximage2?o=9206846&l=v4" scale="0"></a></li> <li> <script type="text/javascript"> gnp_request = {"slug" : "emilys-entourage", "color-set" : 1 , "campaign" 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