Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction
Identifieur interne : 000301 ( Main/Corpus ); précédent : 000300; suivant : 000302Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction
Auteurs : Gusztav Belteki ; Jody Haigh ; Nikolett Kabacs ; Katharina Haigh ; Karen Sison ; Frank Costantini ; Jeff Whitsett ; Susan E. Quaggin ; Andras NagySource :
- Nucleic Acids Research [ 0305-1048 ] ; 2005.
Abstract
Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.
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DOI: 10.1093/nar/gni051
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To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>Gusztav Belteki</name><affiliations><json:string>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</json:string></affiliations></json:item><json:item><name>Jody Haigh</name><affiliations><json:string>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</json:string></affiliations></json:item><json:item><name>Nikolett Kabacs</name><affiliations><json:string>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</json:string></affiliations></json:item><json:item><name>Katharina Haigh</name><affiliations><json:string>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</json:string></affiliations></json:item><json:item><name>Karen Sison</name><affiliations><json:string>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</json:string></affiliations></json:item><json:item><name>Frank Costantini</name><affiliations><json:string>Department of Genetics and Development, College of Physicians and Surgeons, Columbia University New York, NY 10032, USA</json:string></affiliations></json:item><json:item><name>Jeff Whitsett</name><affiliations><json:string>Children's Hospital Medical Center, Division of Pulmonary Biology Cincinnati, OH 45229-3039, USA</json:string></affiliations></json:item><json:item><name>Susan E. 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We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. 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Tel: +1 416 586 3246; Fax: +1 416 586 8588; Email: nagy@mshri.on.ca</affiliation></author></analytic><monogr><title level="j">Nucleic Acids Research</title><title level="j" type="abbrev">Nucl. 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To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.</p></abstract><textClass><keywords scheme="keyword"><list><head>heading</head><item><term>Methods Online</term></item></list></keywords></textClass><textClass><keywords scheme="keyword"><list><head>heading</head><item><term>19</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2005">Published</change><change xml:id="refBibs-istex" who="#ISTEX-API" when="2016-1-2">References added</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/575A726A7C2FF59BC3829CD72A1E03EE4B5A9F41/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="US-ASCII"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article xml:lang="en" article-type="other"><front><journal-meta><journal-id journal-id-type="hwp">nar</journal-id><journal-id journal-id-type="nlm-ta">Nucleic Acids Res</journal-id><journal-id journal-id-type="publisher-id">nar</journal-id><journal-title>Nucleic Acids Research</journal-title><abbrev-journal-title abbrev-type="publisher">Nucl. Acids Res.</abbrev-journal-title><issn pub-type="ppub">0305-1048</issn><issn pub-type="epub">1362-4962</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="other">gni051</article-id><article-id pub-id-type="doi">10.1093/nar/gni051</article-id><article-categories><subj-group subj-group-type="heading"><subject>Methods Online</subject></subj-group><subj-group subj-group-type="hwp-journal-coll"><subject>19</subject></subj-group></article-categories><title-group><article-title>Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Belteki</surname><given-names>Gusztav</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Haigh</surname><given-names>Jody</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kabacs</surname><given-names>Nikolett</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Haigh</surname><given-names>Katharina</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Sison</surname><given-names>Karen</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Costantini</surname><given-names>Frank</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Whitsett</surname><given-names>Jeff</given-names></name><xref rid="AU3">3</xref></contrib><contrib contrib-type="author"><name><surname>Quaggin</surname><given-names>Susan E.</given-names></name><xref rid="AU1">1</xref><xref rid="AU4">4</xref></contrib><contrib contrib-type="author"><name><surname>Nagy</surname><given-names>Andras</given-names></name><xref rid="AU1">1</xref><xref rid="AU5">5</xref><xref rid="COR1">*</xref></contrib><aff id="AU1"><sup>1</sup>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</aff><aff id="AU2"><sup>2</sup>Department of Genetics and Development, College of Physicians and Surgeons, Columbia University New York, NY 10032, USA</aff><aff id="AU3"><sup>3</sup>Children's Hospital Medical Center, Division of Pulmonary Biology Cincinnati, OH 45229-3039, USA</aff><aff id="AU4"><sup>4</sup>St Michael's Hospital Toronto, Ontario, Canada</aff><aff id="AU5"><sup>5</sup>Department of Molecular and Medical Genetics, University of Toronto Toronto, Ontario, Canada M5S 1A8</aff></contrib-group><author-notes><corresp id="COR1"><sup>*</sup>To whom correspondence should be addressed. Tel: +1 416 586 3246; Fax: +1 416 586 8588; Email: <ext-link xlink:href="nagy@mshri.on.ca" ext-link-type="email">nagy@mshri.on.ca</ext-link></corresp></author-notes><pub-date pub-type="ppub"><year>2005</year></pub-date><volume>33</volume><issue>5</issue><fpage>e51</fpage><lpage>e51</lpage><history><date date-type="accepted"><day>01</day><month>3</month><year>2005</year></date><date date-type="received"><day>20</day><month>1</month><year>2005</year></date><date date-type="rev-recd"><day>01</day><month>3</month><year>2005</year></date></history><permissions><copyright-statement>© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact <ext-link xlink:href="journals.permissions@oupjournals.org" ext-link-type="email">journals.permissions@oupjournals.org</ext-link></copyright-statement><copyright-year>2005</copyright-year><license license-type="open-access"><p></p></license></permissions><abstract xml:lang="en"><p>Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (<italic>tet</italic>-<italic>O</italic>)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.</p></abstract><custom-meta-wrap><custom-meta><meta-name>hwp-legacy-fpage</meta-name><meta-value>e51</meta-value></custom-meta><custom-meta><meta-name>hwp-legacy-dochead</meta-name><meta-value>research-article</meta-value></custom-meta></custom-meta-wrap></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction</title></titleInfo><name type="personal"><namePart type="given">Gusztav</namePart><namePart type="family">Belteki</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jody</namePart><namePart type="family">Haigh</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Nikolett</namePart><namePart type="family">Kabacs</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Katharina</namePart><namePart type="family">Haigh</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Karen</namePart><namePart type="family">Sison</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Frank</namePart><namePart type="family">Costantini</namePart><affiliation>Department of Genetics and Development, College of Physicians and Surgeons, Columbia University New York, NY 10032, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jeff</namePart><namePart type="family">Whitsett</namePart><affiliation>Children's Hospital Medical Center, Division of Pulmonary Biology Cincinnati, OH 45229-3039, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Susan E.</namePart><namePart type="family">Quaggin</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><affiliation>St Michael's Hospital Toronto, Ontario, Canada</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Andras</namePart><namePart type="family">Nagy</namePart><affiliation>Samuel Lunenfeld Research Institute, Mount Sinai Hospital 600 University Avenue, Toronto, Ontario, Canada M5G 1X5</affiliation><affiliation>Department of Molecular and Medical Genetics, University of Toronto Toronto, Ontario, Canada M5S 1A8</affiliation><affiliation>To whom correspondence should be addressed. Tel: +1 416 586 3246; Fax: +1 416 586 8588; Email: nagy@mshri.on.ca</affiliation><affiliation>E-mail: nagy@mshri.on.ca</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="other">other</genre><subject><genre>heading</genre><topic>Methods Online</topic></subject><subject><genre>heading</genre><topic>19</topic></subject><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">2005</dateIssued><copyrightDate encoding="w3cdtf">2005</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.</abstract><note type="author-notes">*To whom correspondence should be addressed. Tel: +1 416 586 3246; Fax: +1 416 586 8588; Email: nagy@mshri.on.ca</note><relatedItem type="host"><titleInfo><title>Nucleic Acids Research</title></titleInfo><titleInfo type="abbreviated"><title>Nucl. Acids Res.</title></titleInfo><identifier type="ISSN">0305-1048</identifier><identifier type="eISSN">1362-4962</identifier><identifier type="JournalID">nar</identifier><identifier type="JournalID-hwp">nar</identifier><identifier type="JournalID-nlm-ta">Nucleic Acids Res</identifier><part><date>2005</date><detail type="volume"><caption>vol.</caption><number>33</number></detail><detail type="issue"><caption>no.</caption><number>5</number></detail><extent unit="pages"><start>e51</start><end>e51</end></extent></part></relatedItem><identifier type="istex">575A726A7C2FF59BC3829CD72A1E03EE4B5A9F41</identifier><identifier type="DOI">10.1093/nar/gni051</identifier><accessCondition type="use and reproduction" contentType="Copyright">© The Author 2005. Published by Oxford University Press. 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