Strong and ubiquitous expression of transgenes targeted into the β-actin locus by Cre/lox cassette replacement
Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. Here we present an efficient method for rapi...
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| Veröffentlicht in: | Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2005-08, Vol.42 (4), p.229-235 |
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| creator | Shmerling, Doron Danzer, Claus-Peter Mao, Xiaohong Boisclair, Julie Haffner, Michel Lemaistre, Marianne Schuler, Valerie Kaeslin, Edgar Korn, Reinhard Bürki, Kurt Ledermann, Birgit Kinzel, Bernd Müller, Matthias |
| description | Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. Here we present an efficient method for rapid and reproducible transgenesis using the recombinase mediated cassette exchange (RMCE) (Bouhassira et al.: Blood 90:3332–3344, 1997) procedure. A lox511‐EGFP‐TK/neo‐loxP cassette was placed under the control of the endogenous mouse β‐actin promoter. Heterozygous mice revealed strong and ubiquitous EGFP expression throughout embryogenesis and adulthood. Reproducibly, the same expression pattern was obtained with RMCE when it was used to replace the EGFP‐harboring cassette by ECFP or placental alkaline phosphatase (PLAP) reporter genes (DePrimo et al.: Transgenic Res 5:459–466, 1996). Furthermore, the RMCE procedure proved efficient as well in embryonic stem (ES) cells as directly in zygotes. Our results demonstrate ubiquitous expression of floxed transgenes in the endogenous β‐actin locus and they support the general use of the β‐actin locus for targeted transgenesis. genesis 42:229–235, 2005. © 2005 Wiley‐Liss, Inc. |
| doi_str_mv | 10.1002/gene.20135 |
| format | Article |
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Here we present an efficient method for rapid and reproducible transgenesis using the recombinase mediated cassette exchange (RMCE) (Bouhassira et al.: Blood 90:3332–3344, 1997) procedure. A lox511‐EGFP‐TK/neo‐loxP cassette was placed under the control of the endogenous mouse β‐actin promoter. Heterozygous mice revealed strong and ubiquitous EGFP expression throughout embryogenesis and adulthood. Reproducibly, the same expression pattern was obtained with RMCE when it was used to replace the EGFP‐harboring cassette by ECFP or placental alkaline phosphatase (PLAP) reporter genes (DePrimo et al.: Transgenic Res 5:459–466, 1996). Furthermore, the RMCE procedure proved efficient as well in embryonic stem (ES) cells as directly in zygotes. Our results demonstrate ubiquitous expression of floxed transgenes in the endogenous β‐actin locus and they support the general use of the β‐actin locus for targeted transgenesis. genesis 42:229–235, 2005. © 2005 Wiley‐Liss, Inc.</description><identifier>ISSN: 1526-954X</identifier><identifier>EISSN: 1526-968X</identifier><identifier>DOI: 10.1002/gene.20135</identifier><identifier>PMID: 16028230</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Actins - genetics ; AGRP ; Animals ; Gene Expression ; Gene Targeting - methods ; Genetic Vectors ; Immunohistochemistry ; lox511 ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic - genetics ; mouse ; Oocytes ; Plasmids ; Recombinases - genetics ; Recombinases - metabolism ; RMCE ; Stem Cells ; targeted transgenesis ; Transfection ; Transgenes - physiology</subject><ispartof>Genesis (New York, N.Y. : 2000), 2005-08, Vol.42 (4), p.229-235</ispartof><rights>Copyright © 2005 Wiley‐Liss, Inc.</rights><rights>Copyright 2005 Wiley-Liss, Inc</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3965-87af22e76d2348893e92a28420392ae876ee46c85c3fd62c6cba2738a20c24e33</citedby><cites>FETCH-LOGICAL-c3965-87af22e76d2348893e92a28420392ae876ee46c85c3fd62c6cba2738a20c24e33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fgene.20135$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fgene.20135$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16028230$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shmerling, Doron</creatorcontrib><creatorcontrib>Danzer, Claus-Peter</creatorcontrib><creatorcontrib>Mao, Xiaohong</creatorcontrib><creatorcontrib>Boisclair, Julie</creatorcontrib><creatorcontrib>Haffner, Michel</creatorcontrib><creatorcontrib>Lemaistre, Marianne</creatorcontrib><creatorcontrib>Schuler, Valerie</creatorcontrib><creatorcontrib>Kaeslin, Edgar</creatorcontrib><creatorcontrib>Korn, Reinhard</creatorcontrib><creatorcontrib>Bürki, Kurt</creatorcontrib><creatorcontrib>Ledermann, Birgit</creatorcontrib><creatorcontrib>Kinzel, Bernd</creatorcontrib><creatorcontrib>Müller, Matthias</creatorcontrib><title>Strong and ubiquitous expression of transgenes targeted into the β-actin locus by Cre/lox cassette replacement</title><title>Genesis (New York, N.Y. : 2000)</title><addtitle>Genesis</addtitle><description>Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. 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Danzer, Claus-Peter ; Mao, Xiaohong ; Boisclair, Julie ; Haffner, Michel ; Lemaistre, Marianne ; Schuler, Valerie ; Kaeslin, Edgar ; Korn, Reinhard ; Bürki, Kurt ; Ledermann, Birgit ; Kinzel, Bernd ; Müller, Matthias</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3965-87af22e76d2348893e92a28420392ae876ee46c85c3fd62c6cba2738a20c24e33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Actins - genetics</topic><topic>AGRP</topic><topic>Animals</topic><topic>Gene Expression</topic><topic>Gene Targeting - methods</topic><topic>Genetic Vectors</topic><topic>Immunohistochemistry</topic><topic>lox511</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Mice, Transgenic - genetics</topic><topic>mouse</topic><topic>Oocytes</topic><topic>Plasmids</topic><topic>Recombinases - genetics</topic><topic>Recombinases - metabolism</topic><topic>RMCE</topic><topic>Stem Cells</topic><topic>targeted transgenesis</topic><topic>Transfection</topic><topic>Transgenes - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shmerling, Doron</creatorcontrib><creatorcontrib>Danzer, Claus-Peter</creatorcontrib><creatorcontrib>Mao, Xiaohong</creatorcontrib><creatorcontrib>Boisclair, Julie</creatorcontrib><creatorcontrib>Haffner, Michel</creatorcontrib><creatorcontrib>Lemaistre, Marianne</creatorcontrib><creatorcontrib>Schuler, Valerie</creatorcontrib><creatorcontrib>Kaeslin, Edgar</creatorcontrib><creatorcontrib>Korn, Reinhard</creatorcontrib><creatorcontrib>Bürki, Kurt</creatorcontrib><creatorcontrib>Ledermann, Birgit</creatorcontrib><creatorcontrib>Kinzel, Bernd</creatorcontrib><creatorcontrib>Müller, Matthias</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Genesis (New York, N.Y. : 2000)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shmerling, Doron</au><au>Danzer, Claus-Peter</au><au>Mao, Xiaohong</au><au>Boisclair, Julie</au><au>Haffner, Michel</au><au>Lemaistre, Marianne</au><au>Schuler, Valerie</au><au>Kaeslin, Edgar</au><au>Korn, Reinhard</au><au>Bürki, Kurt</au><au>Ledermann, Birgit</au><au>Kinzel, Bernd</au><au>Müller, Matthias</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Strong and ubiquitous expression of transgenes targeted into the β-actin locus by Cre/lox cassette replacement</atitle><jtitle>Genesis (New York, N.Y. : 2000)</jtitle><addtitle>Genesis</addtitle><date>2005-08</date><risdate>2005</risdate><volume>42</volume><issue>4</issue><spage>229</spage><epage>235</epage><pages>229-235</pages><issn>1526-954X</issn><eissn>1526-968X</eissn><abstract>Conventional approaches to produce transgenic mice recurrently yield unpredictable patterns and levels of transgene expression, a situation calling for the development of new techniques to overcome these drawbacks in the context of overexpression studies. 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| identifier | ISSN: 1526-954X |
| ispartof | Genesis (New York, N.Y. : 2000), 2005-08, Vol.42 (4), p.229-235 |
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| language | eng |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Actins - genetics AGRP Animals Gene Expression Gene Targeting - methods Genetic Vectors Immunohistochemistry lox511 Mice Mice, Inbred BALB C Mice, Transgenic - genetics mouse Oocytes Plasmids Recombinases - genetics Recombinases - metabolism RMCE Stem Cells targeted transgenesis Transfection Transgenes - physiology |
| title | Strong and ubiquitous expression of transgenes targeted into the β-actin locus by Cre/lox cassette replacement |
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