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Creators/Authors contains: "Maverakis, Emanual"

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  1. null (Ed.)
  2. Abstract

    Genetically encoded fluorescent voltage indicators, such as ArcLight, have been used to report action potentials (APs) in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). However, the ArcLight expression, in all cases, relied on a high number of lentiviral vector-mediated random genome integrations (8-12 copy/cell), raising concerns such as gene disruption and alteration of global and local gene expression, as well as loss or silencing of reporter genes after differentiation. Here, we report the use of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 nuclease technique to develop a hiPSC line stably expressing ArcLight from the AAVS1 safe harbor locus. The hiPSC line retained proliferative ability with a growth rate similar to its parental strain. Optical recording with conventional epifluorescence microscopy allowed the detection of APs as early as 21 days postdifferentiation, and could be repeatedly monitored for at least 5 months. Moreover, quantification and analysis of the APs of ArcLight-CMs identified two distinctive subtypes: a group with high frequency of spontaneous APs of small amplitudes that were pacemaker-like CMs and a group with low frequency of automaticity and large amplitudes that resembled the working CMs. Compared with FluoVolt voltage-sensitive dye, although dimmer, the ArcLight reporter exhibited better optical performance in terms of phototoxicity and photostability with comparable sensitivities and signal-to-noise ratios. The hiPSC line with targeted ArcLight engineering design represents a useful tool for studying cardiac development or hiPSC-derived cardiac disease models and drug testing.

    </sec> </div> <a href='#' class='show open-abstract' style='margin-left:10px;'>more »</a> <a href='#' class='hide close-abstract' style='margin-left:10px;'>« less</a> <div class="actions" style="padding-left:10px;"> </div> </div><div class="clearfix"></div> </div> </li> <li> <div class="article item document" itemscope itemtype="http://schema.org/TechArticle"> <div class="item-info"> <div class="title"> <a href="https://par.nsf.gov/biblio/10322588-guidelines-use-interpretation-assays-monitoring-autophagy-edition" itemprop="url"> <span class='span-link' itemprop="name">Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1</span> </a> </div> <div> <strong> <a class="misc external-link" href="https://doi.org/10.1080/15548627.2020.1797280" target="_blank" title="Link to document DOI">https://doi.org/10.1080/15548627.2020.1797280  <span class="fas fa-external-link-alt"></span></a> </strong> </div> <div class="metadata"> <span class="authors"> <span class="author" itemprop="author">Klionsky, Daniel J.</span> <span class="sep">; </span><span class="author" itemprop="author">Abdel-Aziz, Amal Kamal</span> <span class="sep">; </span><span class="author" itemprop="author">Abdelfatah, Sara</span> <span class="sep">; </span><span class="author" itemprop="author">Abdellatif, Mahmoud</span> <span class="sep">; </span><span class="author" itemprop="author">Abdoli, Asghar</span> <span class="sep">; </span><span class="author" itemprop="author">Abel, Steffen</span> <span class="sep">; </span><span class="author" itemprop="author">Abeliovich, Hagai</span> <span class="sep">; </span><span class="author" itemprop="author">Abildgaard, Marie H.</span> <span class="sep">; </span><span class="author" itemprop="author">Abudu, Yakubu Princely</span> <span class="sep">; </span><span class="author" itemprop="author">Acevedo-Arozena, Abraham</span> <span class="sep">; </span><span class="author">et al</span></span> <span class="year">( <time itemprop="datePublished" datetime="2021-01-02">January 2021</time> , Autophagy) </span> </div> <div class="actions" style="padding-left:10px;"> <span class="reader-count"> <a class="misc external-link" href="https://doi.org/10.1080/15548627.2020.1797280" target="_blank" title="Link to document DOI" data-ostiid="10322588"> Full Text Available <span class="fas fa-external-link-alt"></span> </a> </span> </div> </div><div class="clearfix"></div> </div> </li> </ol> <div id="pagination-lower" style="display:none;"> <div class="pull-right" style="line-height: 30px;"> <div class="btn-group pagination nomargin"> <a href="#" class="btn btn-sm btn-default noborderradius" disabled="disabled">«<span class="hidden-xs"> Prev</span></a> <a class="dropdown-toggle btn btn-sm btn-default paging-dropdown disabled hidden-xs noborderradius" href="#" data-toggle="dropdown"> <span class="caret"></span> <span class="sr-only">Select page number</span> </a> <a href="#" class="btn btn-sm btn-default noborderradius" disabled="disabled"><span class="hidden-xs">Next </span>»</a> </div> </div> <div class="clearfix"></div> </div> </div> </div> </div> <!-- / basic search results --> </div> </div> <input type='hidden' id='schtype' value=''/> </div> </div> <footer class="row" id="footer-wrapper"> <div class="footer-content"> <div id="footerOSTI" class=" hidden-print"> <ul> <li><a target="_blank" rel="noreferrer" href="http://www.nsf.gov/policies/">Website Policies</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/about/performance/">Budget and Performance</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/oig/">Inspector General</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/policies/privacy.jsp">Privacy</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/policies/foia.jsp">FOIA</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/od/odi/notice.jsp">No FEAR Act</a> | <a target="_blank" rel="noreferrer" href="http://usa.gov">USA.gov</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/policies/access.jsp">Accessibility</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/policies/nsf_plain_language.jsp">Plain Language</a> | <a target="_blank" rel="noreferrer" href="http://www.nsf.gov/help/contact.jsp">Contact</a> | <a target="_blank" rel="noreferrer" href="https://nsf.gov/help/">Help</a> </li> </ul> The National Science Foundation, 2415 Eisenhower Avenue, Alexandria, Virginia 22314, USA Tel: (703) 292-5111, FIRS: (800) 877-8339 | TDD: (800) 281-8749 </div> </div> </footer> </div> <div id="authorselect" class="modal" tabindex="-1" role="dialog" aria-labelledby="authorselect_label" aria-hidden="true"> <div class="modal-dialog"> <div class="modal-content"> <div class="modal-header"> <button type="button" class="close" data-dismiss="modal" aria-hidden="true">×</button> <div id="authorselect_label">Author Select</div> </div> <form id="authorselect-form" style="margin-bottom: 0px;"> <input type="hidden" name="pg" id="authorselect-pg" value="1" /> <div class="modal-body" id="authorselect_body"> <div class="row"> <div class="col-md-4"> <label for="authorselect-lname">Last Name:</label><br /> <input type="text" name="lname" class="input-sm form-control" id="authorselect-lname" placeholder="Last name" /><br /> </div> <div class="col-md-4"> <label for="authorselect-fname">First Name:</label><br /> <input type="text" name="fname" class="input-sm form-control" id="authorselect-fname" placeholder="First name" /> </div> <div class="col-md-2">  <br /> <a href="#" onclick="$('#authorselect-form').submit(); 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