Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors
Identifieur interne : 000529 ( Main/Corpus ); précédent : 000528; suivant : 000530Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors
Auteurs : Stelios Psarras ; Niki Karagianni ; Christoph Kellendonk ; François Tronche ; François Oic Cosset ; Carol Stocking ; Volker Schirrmacher ; Harald Von Boehmer ; Khashayarsha KhazaieSource :
- The Journal of Gene Medicine [ 1099-498X ] ; 2004-01.
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- KwdEn :
Abstract
Background: Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods: To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase. Results: Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50. Conclusions: The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.
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DOI: 10.1002/jgm.442
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first="Christoph" last="Kellendonk">Christoph Kellendonk</name><affiliation><mods:affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Tronche, Francois" sort="Tronche, Francois" uniqKey="Tronche F" first="François" last="Tronche">François Tronche</name><affiliation><mods:affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Cosset, Francois Oic" sort="Cosset, Francois Oic" uniqKey="Cosset F" first="François Oic" last="Cosset">François Oic Cosset</name><affiliation><mods:affiliation>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</mods:affiliation></affiliation></author><author><name sortKey="Stocking, Carol" sort="Stocking, Carol" uniqKey="Stocking C" first="Carol" last="Stocking">Carol Stocking</name><affiliation><mods:affiliation>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Schirrmacher, Volker" sort="Schirrmacher, Volker" uniqKey="Schirrmacher V" first="Volker" last="Schirrmacher">Volker Schirrmacher</name><affiliation><mods:affiliation>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Boehmer, Harald Von" sort="Boehmer, Harald Von" uniqKey="Boehmer H" first="Harald Von" last="Boehmer">Harald Von Boehmer</name><affiliation><mods:affiliation>INSERM U373, Institut Necker, Paris, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</mods:affiliation></affiliation></author><author><name sortKey="Khazaie, Khashayarsha" sort="Khazaie, Khashayarsha" 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Stelios" uniqKey="Psarras S" first="Stelios" last="Psarras">Stelios Psarras</name><affiliation><mods:affiliation>INSERM U373, Institut Necker, Paris, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Current Address: Allergy Unit, 2nd Pediatric Clinic, University of Athens, Greece.</mods:affiliation></affiliation></author><author><name sortKey="Karagianni, Niki" sort="Karagianni, Niki" uniqKey="Karagianni N" first="Niki" last="Karagianni">Niki Karagianni</name><affiliation><mods:affiliation>INSERM U373, Institut Necker, Paris, France</mods:affiliation></affiliation></author><author><name sortKey="Kellendonk, Christoph" sort="Kellendonk, Christoph" uniqKey="Kellendonk C" first="Christoph" last="Kellendonk">Christoph Kellendonk</name><affiliation><mods:affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Tronche, Francois" sort="Tronche, Francois" uniqKey="Tronche F" first="François" last="Tronche">François Tronche</name><affiliation><mods:affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Cosset, Francois Oic" sort="Cosset, Francois Oic" uniqKey="Cosset F" first="François Oic" last="Cosset">François Oic Cosset</name><affiliation><mods:affiliation>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</mods:affiliation></affiliation></author><author><name sortKey="Stocking, Carol" sort="Stocking, Carol" uniqKey="Stocking C" first="Carol" last="Stocking">Carol Stocking</name><affiliation><mods:affiliation>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Schirrmacher, Volker" sort="Schirrmacher, Volker" uniqKey="Schirrmacher V" first="Volker" last="Schirrmacher">Volker Schirrmacher</name><affiliation><mods:affiliation>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</mods:affiliation></affiliation></author><author><name sortKey="Boehmer, Harald Von" sort="Boehmer, Harald Von" uniqKey="Boehmer H" first="Harald Von" last="Boehmer">Harald Von Boehmer</name><affiliation><mods:affiliation>INSERM U373, Institut Necker, Paris, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</mods:affiliation></affiliation></author><author><name sortKey="Khazaie, Khashayarsha" sort="Khazaie, Khashayarsha" uniqKey="Khazaie K" first="Khashayarsha" last="Khazaie">Khashayarsha Khazaie</name><affiliation><mods:affiliation>INSERM U373, Institut Necker, Paris, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer 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Gene Med.</title><idno type="ISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><imprint><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2004-01">2004-01</date><biblScope unit="volume">6</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="32">32</biblScope><biblScope unit="page" to="42">42</biblScope></imprint><idno type="ISSN">1099-498X</idno></series><idno type="istex">DDFA1B3954E5822528029DF079EA805025138BD8</idno><idno type="DOI">10.1002/jgm.442</idno><idno type="ArticleID">JGM442</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1099-498X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cre recombinase</term><term>Cre.PR fusion</term><term>ES cells</term><term>MESV retroviral vectors</term><term>adenoviral vectors</term><term>gene transfer</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Background: Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods: To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase. Results: Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50. Conclusions: The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.</div></front></TEI><istex><corpusName>wiley</corpusName><author><json:item><name>Stelios Psarras</name><affiliations><json:string>INSERM U373, Institut Necker, Paris, France</json:string><json:string>Current Address: Allergy Unit, 2nd Pediatric Clinic, University of Athens, Greece.</json:string></affiliations></json:item><json:item><name>Niki Karagianni</name><affiliations><json:string>INSERM U373, Institut Necker, Paris, France</json:string></affiliations></json:item><json:item><name>Christoph Kellendonk</name><affiliations><json:string>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</json:string></affiliations></json:item><json:item><name>François Tronche</name><affiliations><json:string>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</json:string></affiliations></json:item><json:item><name>François‐Loic Cosset</name><affiliations><json:string>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</json:string></affiliations></json:item><json:item><name>Carol Stocking</name><affiliations><json:string>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</json:string></affiliations></json:item><json:item><name>Volker Schirrmacher</name><affiliations><json:string>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</json:string></affiliations></json:item><json:item><name>Harald von Boehmer</name><affiliations><json:string>INSERM U373, Institut Necker, Paris, France</json:string><json:string>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</json:string></affiliations></json:item><json:item><name>Khashayarsha Khazaie</name><affiliations><json:string>INSERM U373, Institut Necker, Paris, France</json:string><json:string>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>gene transfer</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>ES cells</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>MESV retroviral vectors</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>adenoviral vectors</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Cre recombinase</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Cre.PR fusion</value></json:item></subject><language><json:string>eng</json:string></language><abstract>Background: Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods: To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase. Results: Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50. Conclusions: The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.</abstract><qualityIndicators><score>7.58</score><pdfVersion>1.3</pdfVersion><pdfPageSize>595 x 859 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>6</keywordCount><abstractCharCount>1516</abstractCharCount><pdfWordCount>6767</pdfWordCount><pdfCharCount>42475</pdfCharCount><pdfPageCount>11</pdfPageCount><abstractWordCount>215</abstractWordCount></qualityIndicators><title>Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title><genre><json:string>article</json:string></genre><host><volume>6</volume><pages><total>11</total><last>42</last><first>32</first></pages><issn><json:string>1099-498X</json:string></issn><issue>1</issue><subject><json:item><value>Research Article</value></json:item></subject><genre></genre><language><json:string>unknown</json:string></language><eissn><json:string>1521-2254</json:string></eissn><title>The Journal of Gene Medicine</title><doi><json:string>10.1002/(ISSN)1521-2254</json:string></doi></host><publicationDate>2004</publicationDate><copyrightDate>2004</copyrightDate><doi><json:string>10.1002/jgm.442</json:string></doi><id>DDFA1B3954E5822528029DF079EA805025138BD8</id><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/DDFA1B3954E5822528029DF079EA805025138BD8/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/DDFA1B3954E5822528029DF079EA805025138BD8/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/DDFA1B3954E5822528029DF079EA805025138BD8/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><availability><p>WILEY</p></availability><date>2004</date></publicationStmt><notesStmt><note>Department of the Army, Breast Cancer Research Idea Award - No. DAMD17‐02‐1‐0361;</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title><author><persName><forename type="first">Stelios</forename><surname>Psarras</surname></persName><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Current Address: Allergy Unit, 2nd Pediatric Clinic, University of Athens, Greece.</affiliation></author><author><persName><forename type="first">Niki</forename><surname>Karagianni</surname></persName><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation></author><author><persName><forename type="first">Christoph</forename><surname>Kellendonk</surname></persName><affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</affiliation></author><author><persName><forename type="first">François</forename><surname>Tronche</surname></persName><affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</affiliation></author><author><persName><forename type="first">François‐Loic</forename><surname>Cosset</surname></persName><affiliation>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</affiliation></author><author><persName><forename type="first">Carol</forename><surname>Stocking</surname></persName><affiliation>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</affiliation></author><author><persName><forename type="first">Volker</forename><surname>Schirrmacher</surname></persName><affiliation>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</affiliation></author><author><persName><forename type="first">Harald von</forename><surname>Boehmer</surname></persName><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</affiliation></author><author><persName><forename type="first">Khashayarsha</forename><surname>Khazaie</surname></persName><note type="correspondence"><p>Correspondence: Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Smith Building 7th Floor, 44 Binney St., Boston, MA 02115, USA.</p></note><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</affiliation></author></analytic><monogr><title level="j">The Journal of Gene Medicine</title><title level="j" type="sub">A cross‐disciplinary journal for research on the science of gene transfer and its clinical applications</title><title level="j" type="abbrev">J. Gene Med.</title><idno type="pISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><idno type="DOI">10.1002/(ISSN)1521-2254</idno><imprint><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2004-01"></date><biblScope unit="volume">6</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="32">32</biblScope><biblScope unit="page" to="42">42</biblScope></imprint></monogr><idno type="istex">DDFA1B3954E5822528029DF079EA805025138BD8</idno><idno type="DOI">10.1002/jgm.442</idno><idno type="ArticleID">JGM442</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2004</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Background: Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods: To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase. Results: Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50. Conclusions: The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>Keywords</head><item><term>gene transfer</term></item><item><term>ES cells</term></item><item><term>MESV retroviral vectors</term></item><item><term>adenoviral vectors</term></item><item><term>Cre recombinase</term></item><item><term>Cre.PR fusion</term></item></list></keywords></textClass><textClass><keywords scheme="Journal Subject"><list><head>article category</head><item><term>Research Article</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2002-03-19">Received</change><change when="2003-05-26">Registration</change><change when="2004-01">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/DDFA1B3954E5822528029DF079EA805025138BD8/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Wiley, elements deleted: body"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="yes"</istex:xmlDeclaration><istex:document><component version="2.0" type="serialArticle" xml:lang="en"><header><publicationMeta level="product"><publisherInfo><publisherName>John Wiley & Sons, Ltd.</publisherName><publisherLoc>Chichester, UK</publisherLoc></publisherInfo><doi registered="yes">10.1002/(ISSN)1521-2254</doi><issn type="print">1099-498X</issn><issn type="electronic">1521-2254</issn><idGroup><id type="product" value="JGM"></id></idGroup><titleGroup><title type="main" xml:lang="en" sort="JOURNAL OF GENE MEDICINE, THE">The Journal of Gene Medicine</title><title type="subtitle">A cross‐disciplinary journal for research on the science of gene transfer and its clinical applications</title><title type="short">J. 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number="5"></count><count type="tableTotal" number="0"></count><count type="referenceTotal" number="54"></count></countGroup><titleGroup><title type="main" xml:lang="en">Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title><title type="short" xml:lang="en">MESV‐Mediated Cre/loxp Modification of ESCells</title></titleGroup><creators><creator xml:id="au1" creatorRole="author" affiliationRef="#af1" currentRef="#curr1"><personName><givenNames>Stelios</givenNames><familyName>Psarras</familyName></personName></creator><creator xml:id="au2" creatorRole="author" affiliationRef="#af1"><personName><givenNames>Niki</givenNames><familyName>Karagianni</familyName></personName></creator><creator xml:id="au3" creatorRole="author" affiliationRef="#af3"><personName><givenNames>Christoph</givenNames><familyName>Kellendonk</familyName></personName></creator><creator xml:id="au4" creatorRole="author" affiliationRef="#af3"><personName><givenNames>François</givenNames><familyName>Tronche</familyName></personName></creator><creator xml:id="au5" creatorRole="author" affiliationRef="#af4"><personName><givenNames>François‐Loic</givenNames><familyName>Cosset</familyName></personName></creator><creator xml:id="au6" creatorRole="author" affiliationRef="#af5"><personName><givenNames>Carol</givenNames><familyName>Stocking</familyName></personName></creator><creator xml:id="au7" creatorRole="author" affiliationRef="#af2"><personName><givenNames>Volker</givenNames><familyName>Schirrmacher</familyName></personName></creator><creator xml:id="au8" creatorRole="author" affiliationRef="#af1 #af6"><personName><givenNames>Harald von</givenNames><familyName>Boehmer</familyName></personName></creator><creator xml:id="au9" creatorRole="author" affiliationRef="#af1 #af6" corresponding="yes"><personName><givenNames>Khashayarsha</givenNames><familyName>Khazaie</familyName></personName><contactDetails><email>Khashayarsha_Khazaie@dfci.harvard.edu</email></contactDetails></creator></creators><affiliationGroup><affiliation xml:id="af1" countryCode="FR" type="organization"><unparsedAffiliation>INSERM U373, Institut Necker, Paris, France</unparsedAffiliation></affiliation><affiliation xml:id="af2" countryCode="DE" type="organization"><unparsedAffiliation>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</unparsedAffiliation></affiliation><affiliation xml:id="af3" countryCode="DE" type="organization"><unparsedAffiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</unparsedAffiliation></affiliation><affiliation xml:id="af4" countryCode="FR" type="organization"><unparsedAffiliation>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</unparsedAffiliation></affiliation><affiliation xml:id="af5" countryCode="DE" type="organization"><unparsedAffiliation>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</unparsedAffiliation></affiliation><affiliation xml:id="af6" countryCode="US" type="organization"><unparsedAffiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</unparsedAffiliation></affiliation><affiliation xml:id="curr1" countryCode="GR"><unparsedAffiliation>Allergy Unit, 2nd Pediatric Clinic, University of Athens, Greece.</unparsedAffiliation></affiliation></affiliationGroup><keywordGroup xml:lang="en" type="author"><keyword xml:id="kwd1">gene transfer</keyword><keyword xml:id="kwd2">ES cells</keyword><keyword xml:id="kwd3">MESV retroviral vectors</keyword><keyword xml:id="kwd4">adenoviral vectors</keyword><keyword xml:id="kwd5">Cre recombinase</keyword><keyword xml:id="kwd6">Cre.PR fusion</keyword></keywordGroup><fundingInfo><fundingAgency>Department of the Army, Breast Cancer Research Idea Award</fundingAgency><fundingNumber>DAMD17‐02‐1‐0361</fundingNumber></fundingInfo><abstractGroup><abstract type="main" xml:lang="en"><title type="main">Abstract</title><section xml:id="abs1-1"><title type="main">Background</title><p>Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells.</p></section><section xml:id="abs1-2"><title type="main">Methods</title><p>To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase.</p></section><section xml:id="abs1-3"><title type="main">Results</title><p>Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50.</p></section><section xml:id="abs1-4"><title type="main">Conclusions</title><p>The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.</p></section></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>MESV‐Mediated Cre/loxp Modification of ESCells</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Gene transfer and genetic modification of embryonic stem cells by Cre‐ and Cre‐PR‐expressing MESV‐based retroviral vectors</title></titleInfo><name type="personal"><namePart type="given">Stelios</namePart><namePart type="family">Psarras</namePart><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Current Address: Allergy Unit, 2nd Pediatric Clinic, University of Athens, Greece.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Niki</namePart><namePart type="family">Karagianni</namePart><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Christoph</namePart><namePart type="family">Kellendonk</namePart><affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">François</namePart><namePart type="family">Tronche</namePart><affiliation>Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">François‐Loic</namePart><namePart type="family">Cosset</namePart><affiliation>Laboratory of Retroviral Vectorology and Gene Therapy, INSERM U412, Lyon, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Carol</namePart><namePart type="family">Stocking</namePart><affiliation>Division for Cellular and Viral Genetics, Heinrich Pette Institute for Experimental Virology and Immunology, University of Hamburg, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Volker</namePart><namePart type="family">Schirrmacher</namePart><affiliation>Division of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Harald von</namePart><namePart type="family">Boehmer</namePart><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Khashayarsha</namePart><namePart type="family">Khazaie</namePart><affiliation>INSERM U373, Institut Necker, Paris, France</affiliation><affiliation>Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Boston, USA</affiliation><description>Correspondence: Department of Cancer Immunology and Aids, Dana Farber Cancer Institute, Smith Building 7th Floor, 44 Binney St., Boston, MA 02115, USA.</description><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article"></genre><originInfo><publisher>John Wiley & Sons, Ltd.</publisher><place><placeTerm type="text">Chichester, UK</placeTerm></place><dateIssued encoding="w3cdtf">2004-01</dateIssued><dateCaptured encoding="w3cdtf">2002-03-19</dateCaptured><dateValid encoding="w3cdtf">2003-05-26</dateValid><copyrightDate encoding="w3cdtf">2004</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType><extent unit="figures">5</extent><extent unit="references">54</extent></physicalDescription><abstract lang="en">Background: Genetic modification of embryonic stem (ES) cells represents a powerful tool for transgenic and developmental experiments. We report that retroviral constructs based on murine embryonal stem cell virus (MESV) can efficiently deliver and express Cre recombinase or a post‐translationally inducible Cre‐Progesterone receptor (Cre.PR) fusion in mouse fibroblasts and ES cells. Methods: To study the vectors a sensitive reporter cell line, 3TZ, was derived from the murine 3T6 fibroblast line that expresses β‐galactosidase only upon Cre‐mediated recombination. This was used together with the ROSA26‐R ES cell Cre‐reporter system or unmodified mouse ES cells as targets of infection. Efficiency of gene transfer was evaluated immunohistochemically by the use of an anti‐Cre polyclonal antibody, and by monitoring the expression of β‐galactosidase. Results: Infection of the 3TZ cells with high titer 718C or 719CP virus revealed efficient gene transduction of constitutive or hormone‐inducible recombinase activity, respectively. The vectors efficiently transduced murine ES cells with Cre, Cre‐PR (fusion of Cre and progesterone receptor) or β‐galactosidase. Cre‐mediated recombination in more than 60% of ROSA26‐R ES cells was achieved when infected by a VSV‐G‐pseudotyped MESV retrovirus at MOI of 50. Conclusions: The MESV‐based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells. Copyright © 2003 John Wiley & Sons, Ltd.</abstract><note type="funding">Department of the Army, Breast Cancer Research Idea Award - No. DAMD17‐02‐1‐0361; </note><subject lang="en"><genre>Keywords</genre><topic>gene transfer</topic><topic>ES cells</topic><topic>MESV retroviral vectors</topic><topic>adenoviral vectors</topic><topic>Cre recombinase</topic><topic>Cre.PR fusion</topic></subject><relatedItem type="host"><titleInfo><title>The Journal of Gene Medicine</title><subTitle>A cross‐disciplinary journal for research on the science of gene transfer and its clinical applications</subTitle></titleInfo><titleInfo type="abbreviated"><title>J. Gene Med.</title></titleInfo><genre type="Journal">journal</genre><subject><genre>article category</genre><topic>Research Article</topic></subject><identifier type="ISSN">1099-498X</identifier><identifier type="eISSN">1521-2254</identifier><identifier type="DOI">10.1002/(ISSN)1521-2254</identifier><identifier type="PublisherID">JGM</identifier><part><date>2004</date><detail type="volume"><caption>vol.</caption><number>6</number></detail><detail type="issue"><caption>no.</caption><number>1</number></detail><extent unit="pages"><start>32</start><end>42</end><total>11</total></extent></part></relatedItem><identifier type="istex">DDFA1B3954E5822528029DF079EA805025138BD8</identifier><identifier type="DOI">10.1002/jgm.442</identifier><identifier type="ArticleID">JGM442</identifier><accessCondition type="use and reproduction" contentType="copyright">Copyright © 2004 John Wiley & Sons, Ltd.</accessCondition><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>John Wiley & Sons, Ltd.</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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