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<meta name="ncbi_db" content="books" /><meta name="ncbi_pdid" content="book-part" /><meta name="ncbi_acc" content="NBK23162" /><meta name="ncbi_domain" content="micad" /><meta name="ncbi_report" content="record" /><meta name="ncbi_type" content="fulltext" /><meta name="ncbi_objectid" content="" /><meta name="ncbi_pcid" content="/NBK23162/" /><meta name="ncbi_pagename" content="Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc - Molecular Imaging and Contrast Agent Database (MICAD) - NCBI Bookshelf" /><meta name="ncbi_bookparttype" content="chapter" /><meta name="ncbi_app" content="bookshelf" />
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<title>Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc - Molecular Imaging and Contrast Agent Database (MICAD) - NCBI Bookshelf</title>
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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="Molecular Imaging and Contrast Agent Database (MICAD) [Internet]" /><meta name="citation_title" content="Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2009/01/05" /><meta name="citation_author" content="Huiming Zhang" /><meta name="citation_pmid" content="20641365" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK23162/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Huiming Zhang" /><meta name="DC.Date" content="2009/01/05" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK23162/" /><meta name="description" content="Adenoviruses (Ads) comprise a non-enveloped icosahedral protein shell (capsid) 70–100 nm in diameter surrounding an inner vial genome core (1). Human Ads consist of 51 distinct serotypes that are classified into six subgroups (species) designated A–F (2). Some serotypes such as serotypes 2 (Ad2) and 5 (Ad5) of species C only induce a mild, non-oncogenic respiratory infection, making them suitable for development of Ad-based vaccines and gene delivery vehicles for systematic administration (1). Their core genome, a linear, double-stranded DNA of ~36 kb, contains five early transcription units (E1A, E1B, E2, E3, and E4), four intermediate units (IVA2, IX, VAI, and VAII), and one later transcription unit (2). The functions in the early transcription regions have been well identified,: E1A activates cell cycles and initiates DNA replication; E1B blocks apoptosis induced by E1A activity; E2 facilitates viral DNA replication; E3 modulates host immune responses; and E4 regulates DNA replication, mRNA transport, and apoptosis (3). As a gene delivery vehicle, the genome core in Ad2 or Ad5 is usually modified with recombinant transcription activators (RTAs), in which foreign DNA with a size of up to 7.5 kb can be inserted in the deleted E1 or E3 regions in the Ads (4). Such genetic modification can incorporate ligands that can recognize specific cellular receptors and/or block the adenoviral naïve receptor (coxsackie-adenovirus receptor (CAR)), so that their tissue specificity is enhanced significantly. For example, the gene for the androgen receptor (AR) regulates the growth of prostate epithelial cells, induces the expression of prostate-specific antigen (PSA), and controls the growth of prostate cancer in the early phase. The AR gene can be introduced to the adenoviral genome to target AR-responsive prostate cancer or metastasis (5). Furthermore, reporter genes such as firefly luciferase (Fluc) (6) can be included for in vivo imaging, or suicide genes such as herpes simplex virus type 1-thymidine kinase (HSV1-tk) can be used for therapeutic applications (7). As an oxygenase (Mw = 62 kDa), Fluc oxidizes the heterocyclic substrate d-luciferin to oxyluciferin injected into the body and emits light in the wavelength range of 400–620 nm (8)." /><meta name="og:title" content="Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc" /><meta name="og:type" content="book" /><meta name="og:description" content="Adenoviruses (Ads) comprise a non-enveloped icosahedral protein shell (capsid) 70–100 nm in diameter surrounding an inner vial genome core (1). Human Ads consist of 51 distinct serotypes that are classified into six subgroups (species) designated A–F (2). Some serotypes such as serotypes 2 (Ad2) and 5 (Ad5) of species C only induce a mild, non-oncogenic respiratory infection, making them suitable for development of Ad-based vaccines and gene delivery vehicles for systematic administration (1). Their core genome, a linear, double-stranded DNA of ~36 kb, contains five early transcription units (E1A, E1B, E2, E3, and E4), four intermediate units (IVA2, IX, VAI, and VAII), and one later transcription unit (2). The functions in the early transcription regions have been well identified,: E1A activates cell cycles and initiates DNA replication; E1B blocks apoptosis induced by E1A activity; E2 facilitates viral DNA replication; E3 modulates host immune responses; and E4 regulates DNA replication, mRNA transport, and apoptosis (3). As a gene delivery vehicle, the genome core in Ad2 or Ad5 is usually modified with recombinant transcription activators (RTAs), in which foreign DNA with a size of up to 7.5 kb can be inserted in the deleted E1 or E3 regions in the Ads (4). Such genetic modification can incorporate ligands that can recognize specific cellular receptors and/or block the adenoviral naïve receptor (coxsackie-adenovirus receptor (CAR)), so that their tissue specificity is enhanced significantly. For example, the gene for the androgen receptor (AR) regulates the growth of prostate epithelial cells, induces the expression of prostate-specific antigen (PSA), and controls the growth of prostate cancer in the early phase. The AR gene can be introduced to the adenoviral genome to target AR-responsive prostate cancer or metastasis (5). Furthermore, reporter genes such as firefly luciferase (Fluc) (6) can be included for in vivo imaging, or suicide genes such as herpes simplex virus type 1-thymidine kinase (HSV1-tk) can be used for therapeutic applications (7). As an oxygenase (Mw = 62 kDa), Fluc oxidizes the heterocyclic substrate d-luciferin to oxyluciferin injected into the body and emits light in the wavelength range of 400–620 nm (8)." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK23162/" /><meta name="og:site_name" content="NCBI Bookshelf" /><meta name="og:image" content="https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" /><meta name="twitter:card" content="summary" /><meta name="twitter:site" content="@ncbibooks" /><meta name="bk-non-canon-loc" content="/books/n/micad/AdTSTAFluc/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK23162/" /><link rel="stylesheet" href="/corehtml/pmc/css/figpopup.css" type="text/css" media="screen" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books.min.css" type="text/css" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books_print.min.css" type="text/css" media="print" /><style type="text/css">p a.figpopup{display:inline !important} .bk_tt {font-family: monospace} .first-line-outdent .bk_ref {display: inline} .body-content h2, .body-content .h2 {border-bottom: 1px solid #97B0C8} .body-content h2.inline {border-bottom: none} a.page-toc-label , .jig-ncbismoothscroll a {text-decoration:none;border:0 !important} .temp-labeled-list .graphic {display:inline-block !important} .temp-labeled-list img{width:100%}</style><script type="text/javascript" src="/corehtml/pmc/js/jquery.hoverIntent.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/common.min.js?_=3.18"> </script><script type="text/javascript" src="/corehtml/pmc/js/large-obj-scrollbars.min.js"> </script><script type="text/javascript">window.name="mainwindow";</script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/book-toc.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/books.min.js"> </script><meta name="book-collection" content="NONE" />
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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/micad/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" alt="Cover of Molecular Imaging and Contrast Agent Database (MICAD)" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Molecular Imaging and Contrast Agent Database (MICAD) [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK23162_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK23162_dtls__"><div>Bethesda (MD): <a href="https://www.ncbi.nlm.nih.gov/" ref="pagearea=page-banner&targetsite=external&targetcat=link&targettype=publisher">National Center for Biotechnology Information (US)</a>; 2004-2013.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/micad/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/micad/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/sosluciferin/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/micad/GLuc/" title="Next page in this title">Next ></a></div></div></div></div></div>
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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK23162_"><span class="title" itemprop="name"><i>Ad5</i>-(PSE-BC)-(GAL4-(VP16)<sub>2</sub>)-(GAL4)<sub>5</sub>-Fluc </span></h1><div itemprop="alternativeHeadline" class="subtitle whole_rhythm">AdTSTA-FL</div><p class="contrib-group"><span itemprop="author">Huiming Zhang</span>, PhD.</p><a data-jig="ncbitoggler" href="#__NBK23162_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK23162_ai__"><div class="contrib half_rhythm"><span itemprop="author">Huiming Zhang</span>, PhD<div class="affiliation small">
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National Center for Biotechnology Information, NLM, NIH, Bethesda, MD,
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<span class="before-email-separator"></span><span class="email-label">Email: </span><a href="mailto:dev@null" data-email="vog.hin.mln.ibcn@dacim" class="oemail">vog.hin.mln.ibcn@dacim</a>
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</div></div></div><p class="small">Created: <span itemprop="datePublished">December 5, 2008</span>; Last Update: <span itemprop="dateModified">January 5, 2009</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="AdTSTAFluc.T1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK23162/table/AdTSTAFluc.T1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__AdTSTAFluc.T1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Chemical name:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><i>Ad5</i>-(PSE-BC)-(GAL4-(VP16)<sub>2</sub>)-(GAL4)<sub>5</sub>-Fluc</td><td rowspan="9" colspan="1" style="text-align:left;vertical-align:middle;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Abbreviated name:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">AdTSTA-FL</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Synonym:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Agent category:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Protein (Virus)</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Target:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Androgen receptor</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Target category:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Receptor</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Method of detection:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Optical imaging</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Source of signal/contrast:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Luciferin</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Activation:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Yes</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Studies:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
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<ul class="simple-list"><li class="half_rhythm"><div>
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<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" />
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<i>In vitro</i>
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</div></li></ul>
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<ul class="simple-list"><li class="half_rhythm"><div>
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<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" /> Rodents
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</div></li></ul>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">No structure is currently available in <a href="http://pubchem.ncbi.nlm.nih.gov" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubChem</a>.</td></tr></tbody></table></div></div><div id="AdTSTAFluc.Background"><h2 id="_AdTSTAFluc_Background_">Background</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=AdTSTA-FL" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Adenoviruses (Ads) comprise a non-enveloped icosahedral protein shell (capsid) 70–100 nm in diameter surrounding an inner vial genome core (<a class="bk_pop" href="#AdTSTAFluc.REF.1">1</a>). Human Ads consist of 51 distinct serotypes that are classified into six subgroups (species) designated A–F (<a class="bk_pop" href="#AdTSTAFluc.REF.2">2</a>). Some serotypes such as serotypes 2 (Ad2) and 5 (Ad5) of species C only induce a mild, non-oncogenic respiratory infection, making them suitable for development of Ad-based vaccines and gene delivery vehicles for systematic administration (<a class="bk_pop" href="#AdTSTAFluc.REF.1">1</a>). Their core genome, a linear, double-stranded DNA of ~36 kb, contains five early transcription units (E1A, E1B, E2, E3, and E4), four intermediate units (IVA2, IX, VAI, and VAII), and one later transcription unit (<a class="bk_pop" href="#AdTSTAFluc.REF.2">2</a>). The functions in the early transcription regions have been well identified,: E1A activates cell cycles and initiates DNA replication; E1B blocks apoptosis induced by E1A activity; E2 facilitates viral DNA replication; E3 modulates host immune responses; and E4 regulates DNA replication, mRNA transport, and apoptosis (<a class="bk_pop" href="#AdTSTAFluc.REF.3">3</a>). As a gene delivery vehicle, the genome core in Ad2 or Ad5 is usually modified with recombinant transcription activators (RTAs), in which foreign DNA with a size of up to 7.5 kb can be inserted in the deleted E1 or E3 regions in the Ads (<a class="bk_pop" href="#AdTSTAFluc.REF.4">4</a>). Such genetic modification can incorporate ligands that can recognize specific cellular receptors and/or block the adenoviral naïve receptor (coxsackie-adenovirus receptor (CAR)), so that their tissue specificity is enhanced significantly. For example, the gene for the androgen receptor (AR) regulates the growth of prostate epithelial cells, induces the expression of prostate-specific antigen (PSA), and controls the growth of prostate cancer in the early phase. The AR gene can be introduced to the adenoviral genome to target AR-responsive prostate cancer or metastasis (<a class="bk_pop" href="#AdTSTAFluc.REF.5">5</a>). Furthermore, reporter genes such as firefly luciferase (Fluc) (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>) can be included for <i>in vivo</i> imaging, or suicide genes such as herpes simplex virus type 1-thymidine kinase (HSV1-tk) can be used for therapeutic applications (<a class="bk_pop" href="#AdTSTAFluc.REF.7">7</a>). As an oxygenase (Mw = 62 kDa), Fluc oxidizes the heterocyclic substrate <span class="small-caps">d</span>-luciferin to oxyluciferin injected into the body and emits light in the wavelength range of 400–620 nm (<a class="bk_pop" href="#AdTSTAFluc.REF.8">8</a>).</p><p>The transcriptional activity of adenoviral genes can be amplified several orders of magnitude with the use of the GAL4-VP2 fusion protein, where GAL4 is a DNA-binding domain (147 amino acids) and VP2 is a two-tandem repeat of the herpes simplex virus VP16 acidic activation domain (78 amino acids) (<a class="bk_pop" href="#AdTSTAFluc.REF.9">9</a>). The two-step transcriptional amplification (TSTA) system is a novel recombinant transcription activation approach for designing a PSA promoter/enhancer, which can provide as high as ~800-fold enhancement in transcription activity compared with its conventional analog (<a class="bk_pop" href="#AdTSTAFluc.REF.10">10</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.11">11</a>). The TSTA system is composed of two parts in general; i.e., an activator BC-VP2 that contains a PSA promoter with a duplicated ARE4 enhancer core (each core has four binding sites for AR) to control the expression of the chimeric protein GAL4-VP2 and provide cell-specific binding, and the reporter (GAL4)<sub>5</sub>-Fluc (G5-FL), which contains a reporter with five GAL4 repeats to generate a GAL4-responsive Fluc signal (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>). The two parts are linked in a head-to-head orientation and are inserted into the E1 region of Ad5 to produce <i>Ad5</i>-(PSE-BC)-(GAL4-(VP16)<sub>2</sub>)-(GAL4)<sub>5</sub>-Fluc (AdTSTA-FL), an optical agent for imaging AR-responsive PSA. Such a construct can provide ~50-fold amplification (<a class="bk_pop" href="#AdTSTAFluc.REF.11">11</a>) <i>via</i> two steps: the PSA regulates the expression of the potent transcription activator GAL4-VP2, which in turn activates a GAL4-responsive reporter Fluc. AdTSTA-FL can function effectively as a gene delivery vehicle and as a lymphotropic agent in prostate cancer and metastasis <i>in vivo</i> (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.12">12</a>).</p></div><div id="AdTSTAFluc.Synthesis"><h2 id="_AdTSTAFluc_Synthesis_">Synthesis</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20AdTSTA-FL)+AND+synthesis%0D%0A" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>The preparation of AdTSTA-FL was conducted in several steps (<a class="bk_pop" href="#AdTSTAFluc.REF.5">5</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.10">10</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.11">11</a>, <a class="bk_pop" href="#AdTSTAFluc.REF.13">13</a>). The commercial plasmid pBS II SK+ was used as the starting baseline construct pPSE (<a class="bk_pop" href="#AdTSTAFluc.REF.11">11</a>). A <i>Not</i>I-flanked PSA enhancer/promoter expression cassette derived from plasmid PSAR2.4k-PCPSA-P-Lux was inserted into the pPSE to produce PSE. The synthetic ARE4 element that contained four key ARE elements was used to generate a duplicated enhancer core (<a class="bk_pop" href="#AdTSTAFluc.REF.13">13</a>). The insertion of this duplicated core (C) in the PSE replaced the sequence between -3935 and -2855 in the PSE to produce a plasmid vector PBC. Then a GAL-VP2 fragment was inserted into PBC followed by creation of a unique <i>Not</i>I site. A <i>Not</i>I fragment, which contained both the PBC promoter and GAL-VP2 gene, was excised from the obtained plasmid PBC-VP2 and cloned into the <i>Not</i>I site of G5-FL to generate the vector PBC-VP2G5-FL (<a class="bk_pop" href="#AdTSTAFluc.REF.10">10</a>). A <i>Sal</i>I-<i>Not</i>l fragment containing the PBC-VP2G5-FL fragment was derived from the resulting PBC-VP2G5-FL and excised by <i>Not</i>I and partial <i>Sal</i>I digestion (<a class="bk_pop" href="#AdTSTAFluc.REF.5">5</a>). The produced expression cassette PBC-VP2G5-FL was inserted into the <i>Sal</i>I-<i>Not</i>I site of the commercial vector pShuttle, which was then incorporated into the commercial Ad vector AdEasy through homologous recombination to produce AdTSTA-FL (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>). The virus was propagated in 293 cells, purified on a CsCl gradient, and titered with plaque assays on 293 monolayers.</p></div><div id="AdTSTAFluc.In_Vitro_Studies_Tes"><h2 id="_AdTSTAFluc_In_Vitro_Studies_Tes_"><i>In Vitro</i> Studies: Testing in Cells and Tissues</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20AdTSTA-FL)+AND+%22in+vitro%22" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Zhang et al. examined the activity of AdTSTA-FL <i>in vitro</i> (<a class="bk_pop" href="#AdTSTAFluc.REF.5">5</a>). Androgen-dependent prostate cancer cells (LNCaP) were infected with AdTSTA-FL and incubated up to 48 h. The cells were harvested and lysed for assaying activity. GAL4-VP2 and Fluc levels were enhanced significantly in the presence of the synthetic androgen agonist R1881, with a maximum enhancement at 48 h after transcription. Similar studies were conducted in other cells, including MCF7 cells (androgen-expressing breast cancer cells) and PC3 (androgen-negative prostate cancer cells), but only a low basal level of TSTA expression was observed and did not respond to R1881. Thus the AdTSTA-FL responded only to androgen-dependent prostate cancer cells. AdTSTA-FL displayed 10-fold more ligand-induced Fluc activity than the control, AdCMV-FL.</p><p>Sato et al. examined the specificity of AdTSTA-FL <i>in vitro</i> on the basis of the measurement of infectivity (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>). A variety of cells were used in the study, including LNCaP and LPAC-4 cells, H157 and A549 cells (lung cancer), MCF7 cells, and HepG2 cells (liver cancer). The infectivity in these cells with respect to that in the Hela cells (cervical carcinoma) was evaluated on the basis of their viral DNA uptake, which was 1.7 for LNCaP, 1.6 for H157, 1.5 for MCF7, and 1.1 for LAPC-4 and HepG2. Then the activity of AdTSTA-FL was evaluated in several prostate cancer cell lines, including two androgen-responsive cells (LNCaP and LPAC-4) and two AR-negative cells (DU145 and PC3). With a dose of one plaque-forming unit (pfu, an infectious unit) per cell (1 multiplicity of infection (m.o.i)), the normalized Fluc activity was 4.4-fold lower in LPAC-4 than that in LNCaP, whereas the AR-negative cells exhibited negative results (i.e., ~500-fold lower activity). The luciferase activity in other cancer cells was also measured and normalized to the activity measured for LNCaP, and all had much lower activity: a reduction of 2.9-fold was found in A549, 12-fold in H157, 45-fold in HepG2, 60-fold in Hela, and 200-fold in MCF7.</p><p>Burton et al. assessed the accumulation of AdTSTA-FL in lymph nodes <i>ex vivo</i> (<a class="bk_pop" href="#AdTSTAFluc.REF.12">12</a>). Mice (<i>n</i> = 3) were injected with 1 × 10<sup>8</sup> pfu AdTSTA-FL in 20 μl phosphate-buffered saline at their forepaws. Their draining lymph nodes (the axillary and brachial nodes) were dissected 24 h after injection. The nodal capsules were disrupted for extraction of the adenoviral particles. The transduction luciferase activity was found in ipsilateral axillary and brachial lymph nodes but not in the contralateral nodes. The number of viral genomes transported to and retained in the brachial node was ~60,000 copies, which was equivalent to ~0.1% delivery efficiency. Burton et al. also examined the transverse of AdTSTA-FL to regional lymph nodes in the mouse prostate <i>Pten</i> tumor suppressor gene knockout model (<i>Pten</i><sup>-/-</sup>). Ten-week-old <i>Pten</i><sup>-/-</sup> mice with enlarged carcinoma prostate glands were injected with 1 × 10<sup>8</sup> pfu AdTSTA-FL on their dorsolateral lobe. One hour later, ~1% of injected AdTSTA-FL was found in the periaortic lumbar nodes, with a smaller amount in more distant mesenteric and renal lymph nodes.</p></div><div id="AdTSTAFluc.Animal_Studies"><h2 id="_AdTSTAFluc_Animal_Studies_">Animal Studies</h2><div id="AdTSTAFluc.Rodents"><h3>Rodents</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20AdTSTA-FL)%20+AND++rodentia" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Sato et al. investigated the specificity of AdTSTA-FL <i>in vivo</i> (<a class="bk_pop" href="#AdTSTAFluc.REF.6">6</a>). SCID mice (<i>n</i> = 4, ~25 g) were implanted with LAPC-4 xenografts, and the tumors were allowed to grow for 3 weeks. AdTSTA-FL at a dose of 10<sup>7</sup> pfu in 10 μl was injected into each tumor. A cooled-charged-coupled device (CCD) camera was used to image the <i>in vivo</i> expression over a 22-d period. For each imaging session, the mice were intravenously injected with 150 mg/kg <span class="small-caps">d</span>-luciferin (200 μl). A robust signal was observed 4 d after injection of AdTSTA-FL. Compared with the injection of the control AdCMV-FL, the average activity was 110-fold higher. As a control, naïve mice were used for the study. A low signal appeared only in the lung 22 d after intravenous injection of AdTSTA-FL. The kinetics of the expression after intratumoral injection of AdTSTA-FL was examined with sequential images between day 5 and day 13. The AdTSTA-FL displayed 50- to 100-fold higher levels of Fluc activity than the AdCMV-FL, and both gradually decayed from day 7.</p><p>Burton et al. examined the movement of AdTSTA-FL to lymph nodes <i>in vivo</i> (<a class="bk_pop" href="#AdTSTAFluc.REF.12">12</a>). SCID/Beige (natural killer–deficient) mice were implanted with LAPC-9-VEGF-C tumor cells in the right shoulder, and the tumors were allowed to grow to 1.5 cm in diameter. Tumor-bearing mice and naïve mice were injected with 1 × 10<sup>8</sup> pfu AdTSTA-FL in 20 μl phosphate-buffered saline in their forepaw and imaged with a CCD camera 4 d later. In each imaging session, the mice received 150 mg/kg <span class="small-caps">d</span>-luciferin intraperitoneally 20 min before imaging. A clear Fluc signal was observed in the axillary region of the tumor-bearing mice but not in the naïve mice. Burton et al. further assessed the lymph node metastases in nodal lesions of various sizes (<a class="bk_pop" href="#AdTSTAFluc.REF.12">12</a>). Mice (<i>n</i> = 6) bearing 1-cm LAPC-9-VEGF-C-GFP-RL tumors were used for the study of macroscopic nodal lesions. A strong signal was observed in the ipsilateral axilla of mice with an injection of 1 × 10<sup>8</sup> pfu AdTSTA-FL in both paws. Mice with orthotopic implanted LAPC-9-VEGF-C-GFP-RL tumors were used to examine the extensive peritumoral lymphatics. Fifteen days after tumor implantation, the mice were injected with 1 × 10<sup>8</sup> pfu AdTSTA-FL in both hind paws. An observable luciferase signal was found in the inguinal and periaortic region 4 d later. The regional lymph nodes were excised for histological analysis after imaging, which further confirmed the metastasis lesion (~250 μm) of tumors present in the lymph nodes. As a control, luciferase tagged Ads that used cytomegalovirus (CMV) as promoter (AdCMV-FL) were injected in the forepaw of tumor-bearing mouse. The bioluminescence images demonstrated high nonspecific signal at the site of injection (paw) and in the liver caused by the systemic circulation.</p></div><div id="AdTSTAFluc.Other_NonPrimate_Mam"><h3>Other Non-Primate Mammals</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&db=pubmed&details_term=%22%20SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20%28dog%20OR%20rabbit%20OR%20pig%20OR%20sheep%29" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="AdTSTAFluc.NonHuman_Primates"><h3>Non-Human Primates</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20AdTSTA-FL)%20+AND+(primate+non+human)" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div></div><div id="AdTSTAFluc.Human_Studies"><h2 id="_AdTSTAFluc_Human_Studies_">Human Studies</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20AdTSTA-FL)%20+AND+human" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p><div id="AdTSTAFluc.NIH_Support"><h3>NIH Support</h3><p>CA 82214, CA 92131, CA 101904, CA 92865, GM 08652</p></div></div><div id="AdTSTAFluc.references"><h2 id="_AdTSTAFluc_references_">References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.1">Douglas J.T. Adenoviral vectors for gene therapy. <span><span class="ref-journal">Mol Biotechnol. </span>2007;<span class="ref-vol">36</span>(1):71–80.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/17827541" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 17827541</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.2">Majhen D. , Ambriovic-Ristov A. Adenoviral vectors--how to use them in cancer gene therapy? <span><span class="ref-journal">Virus Res. </span>2006;<span class="ref-vol">119</span>(2):121–33.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16533542" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16533542</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.3">Kuhlmann K.F. , Gouma D.J. , Wesseling J.G. Adenoviral gene therapy for pancreatic cancer: where do we stand? <span><span class="ref-journal">Dig Surg. </span>2008;<span class="ref-vol">25</span>(4):278–92.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/18635930" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18635930</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.4">Nettelbeck D.M. , Jerome V. , Muller R. Gene therapy: designer promoters for tumour targeting. <span><span class="ref-journal">Trends Genet. </span>2000;<span class="ref-vol">16</span>(4):174–81.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10729833" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10729833</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.5">Zhang L. , Johnson M. , Le K.H. , Sato M. , Ilagan R. , Iyer M. , Gambhir S.S. , Wu L. , Carey M. Interrogating androgen receptor function in recurrent prostate cancer. <span><span class="ref-journal">Cancer Res. </span>2003;<span class="ref-vol">63</span>(15):4552–60.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12907631" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12907631</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.6">Sato M. , Johnson M. , Zhang L. , Zhang B. , Le K. , Gambhir S.S. , Carey M. , Wu L. Optimization of adenoviral vectors to direct highly amplified prostate-specific expression for imaging and gene therapy. <span><span class="ref-journal">Mol Ther. </span>2003;<span class="ref-vol">8</span>(5):726–37.</span> [<a href="/pmc/articles/PMC2820502/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2820502</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/14599805" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 14599805</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.7">Johnson M. , Sato M. , Burton J. , Gambhir S.S. , Carey M. , Wu L. Micro-PET/CT monitoring of herpes thymidine kinase suicide gene therapy in a prostate cancer xenograft: the advantage of a cell-specific transcriptional targeting approach. <span><span class="ref-journal">Mol Imaging. </span>2005;<span class="ref-vol">4</span>(4):463–72.</span> [<a href="/pmc/articles/PMC2835410/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2835410</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/16285908" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16285908</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.8">Paulmurugan R. , Gambhir S.S. Firefly luciferase enzyme fragment complementation for imaging in cells and living animals. <span><span class="ref-journal">Anal Chem. </span>2005;<span class="ref-vol">77</span>(5):1295–302.</span> [<a href="/pmc/articles/PMC4154832/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC4154832</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/15732910" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15732910</span></a>]</div></dd><dt>9.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.9">Sadowski I. , Ma J. , Triezenberg S. , Ptashne M. GAL4-VP16 is an unusually potent transcriptional activator. <span><span class="ref-journal">Nature. </span>1988;<span class="ref-vol">335</span>(6190):563–4.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3047590" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 3047590</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.10">Zhang L. , Adams J.Y. , Billick E. , Ilagan R. , Iyer M. , Le K. , Smallwood A. , Gambhir S.S. , Carey M. , Wu L. Molecular engineering of a two-step transcription amplification (TSTA) system for transgene delivery in prostate cancer. <span><span class="ref-journal">Mol Ther. </span>2002;<span class="ref-vol">5</span>(3):223–32.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11863411" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11863411</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.11">Iyer M. , Wu L. , Carey M. , Wang Y. , Smallwood A. , Gambhir S.S. Two-step transcriptional amplification as a method for imaging reporter gene expression using weak promoters. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>2001;<span class="ref-vol">98</span>(25):14595–600.</span> [<a href="/pmc/articles/PMC64727/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC64727</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/11734653" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11734653</span></a>]</div></dd><dt>12.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.12">Burton J.B. , Johnson M. , Sato M. , Koh S.B. , Mulholland D.J. , Stout D. , Chatziioannou A.F. , Phelps M.E. , Wu H. , Wu L. Adenovirus-mediated gene expression imaging to directly detect sentinel lymph node metastasis of prostate cancer. <span><span class="ref-journal">Nat Med. </span>2008;<span class="ref-vol">14</span>(8):882–8.</span> [<a href="/pmc/articles/PMC2811163/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2811163</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18622403" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18622403</span></a>]</div></dd><dt>13.</dt><dd><div class="bk_ref" id="AdTSTAFluc.REF.13">Wu L. , Matherly J. , Smallwood A. , Adams J.Y. , Billick E. , Belldegrun A. , Carey M. Chimeric PSA enhancers exhibit augmented activity in prostate cancer gene therapy vectors. <span><span class="ref-journal">Gene Ther. </span>2001;<span class="ref-vol">8</span>(18):1416–26.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11571582" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11571582</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
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<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK23162/?report=reader">PubReader</a></li><li><a href="/books/NBK23162/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK23162" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK23162" style="display:none" title="Cite this Page"><div class="bk_tt">Zhang H. Ad5-(PSE-BC)-(GAL4-(VP16)2)-(GAL4)5-Fluc. 2008 Dec 5 [Updated 2009 Jan 5]. In: Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. <span class="bk_cite_avail"></span></div></div></li><li><a href="/books/NBK23162/pdf/Bookshelf_NBK23162.pdf">PDF version of this page</a> (142K)</li><li><a href="/books/n/micad/toc/bin/micad.csv">MICAD summary (CSV file)</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#AdTSTAFluc.Background" ref="log$=inpage&link_id=inpage">Background</a></li><li><a href="#AdTSTAFluc.Synthesis" ref="log$=inpage&link_id=inpage">Synthesis</a></li><li><a href="#AdTSTAFluc.In_Vitro_Studies_Tes" ref="log$=inpage&link_id=inpage"><i>In Vitro</i> Studies: Testing in Cells and Tissues</a></li><li><a href="#AdTSTAFluc.Animal_Studies" ref="log$=inpage&link_id=inpage">Animal Studies</a></li><li><a href="#AdTSTAFluc.Human_Studies" ref="log$=inpage&link_id=inpage">Human Studies</a></li><li><a href="#AdTSTAFluc.references" ref="log$=inpage&link_id=inpage">References</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Search MICAD</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="source-application" id="Shutter"></a></div><div class="portlet_content"><form xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" name="frmSearch" method="get" action="/books/NBK5330/" id="frmSearch"><script type="text/javascript" src="/corehtml/pmc//js/bookshelf/micad.js">/**/</script><label class="offscreen_noflow" for="txtfield">Search term</label><input id="txtfield" type="text" name="f1_term" size="22" onKeyPress="KeyPress('micad',event,'/books/NBK5330/','')" /><button name="f1_search" type="submit">Go</button><button onclick="this.form.reset();" type="reset">Clear</button><p><b>Limit my Search:</b></p><div class="clearfix"><label for="detection">Method of detection:</label><div class="right"><select name="detection" id="detection" style="width:200px"><option value="" selected="selected">Any</option><option value="(MRI OR "Magnetic resonance imaging" OR MRS)">MRI</option><option value="Multimodal">Multimodal imaging</option><option value="Optical">Optical imaging</option><option value="PET">PET</option><option value="Photoacoustic">Photoacoustic imaging</option><option value="(SPECT OR planar)">SPECT</option><option value="Ultrasound">Ultrasound</option><option value="(x-ray OR ct)">X-ray, CT</option></select></div></div><div class="clearfix"><label for="signal">Source of signal/contrast:</label><div class="right"><select name="signal" id="signal" style="width:200px"><option value="" selected="selected">Any</option><optgroup label="MRI agents"><option value="(Copper OR Cu)">Copper</option><option value="(Europium OR Eu3+)">Europium</option><option value="(Fluorine OR 19F)">Fluorine</option><option value="(Gadolinium OR Gd3+)">Gadolinium</option><option value=""Hyperpolarized 13C"">Hyperpolarized 13C</option><option value=""Iron oxide"">Iron oxide</option><option value=""Nitroxide radicals"">Nitroxide radicals</option><option value="(Oxygen OR 17O)">Oxygen</option><option value="Thulium">Thulium</option></optgroup><optgroup label="Multimodal agents"><option value="((Gadolinium OR Gd3+) AND Optical)">Gadolinium and optical</option><option value="((Gadolinium OR Gd3+) AND (Gold OR Au))">Gadolinium and Gold</option><option value="("Iron oxide" AND (64Cu OR 124I OR 111In))">Iron oxide and
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/><label for="__micad_btn_6">Any</label></div></form><form xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" name="frmGo" method="get" action="javascript:alert('frmGo:_@action_was_not_set')" id="frmGo"><input name="term" value="." type="hidden" /></form></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=1505282" ref="log$=recordlinks">PMC</a><div class="brieflinkpop offscreen_noflow">PubMed Central citations</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pubmed&DbFrom=books&Cmd=Link&LinkName=books_pubmed_refs&IdsFromResult=1505282" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/20641191" ref="ordinalpos=1&linkpos=1&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Ad5-(PSE-BC)-(GAL4-(VP16)(2))-(GAL4)(5)-sr39tk.</a><span class="source">[Molecular Imaging and Contrast...]</span><div class="brieflinkpop offscreen_noflow"><span 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xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> HSV1-TK/GFP/Fluc.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Zhang H. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Molecular Imaging and Contrast Agent Database (MICAD). 2004</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/14599805" ref="ordinalpos=1&linkpos=3&log$=relatedarticles&logdbfrom=pubmed">Optimization of adenoviral vectors to direct highly amplified prostate-specific expression for imaging and gene therapy.</a><span class="source">[Mol Ther. 2003]</span><div class="brieflinkpop offscreen_noflow">Optimization of adenoviral vectors to direct highly amplified prostate-specific expression for imaging and gene therapy.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Sato M, Johnson M, Zhang L, Zhang B, Le K, Gambhir SS, Carey M, Wu L. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Mol Ther. 2003 Nov; 8(5):726-37. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/11571582" ref="ordinalpos=1&linkpos=4&log$=relatedarticles&logdbfrom=pubmed">Chimeric PSA enhancers exhibit augmented activity in prostate cancer gene therapy vectors.</a><span class="source">[Gene Ther. 2001]</span><div class="brieflinkpop offscreen_noflow">Chimeric PSA enhancers exhibit augmented activity in prostate cancer gene therapy vectors.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Wu L, Matherly J, Smallwood A, Adams JY, Billick E, Belldegrun A, Carey M. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Gene Ther. 2001 Sep; 8(18):1416-26. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/20641681" ref="ordinalpos=1&linkpos=5&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> NFluc-FHA2-Aktpep-CFluc.</a><span class="source">[Molecular Imaging and Contrast...]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> NFluc-FHA2-Aktpep-CFluc.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Zhang H. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Molecular Imaging and Contrast Agent Database (MICAD). 2004</em></div></div></li></ul><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed_reviews&uid=20641365" ref="ordinalpos=1&log$=relatedreviews_seeall&logdbfrom=pubmed">See reviews...</a><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=20641365" ref="ordinalpos=1&log$=relatedarticles_seeall&logdbfrom=pubmed">See all...</a></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Recent Activity</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="recent_activity" id="Shutter"></a></div><div class="portlet_content"><div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" 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