Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging
A series of conditional reporter mouse lines were established in which specific organelles were labeled with fluorescent proteins. Subcellular localization and intensity of 28 fluorescent fusion‐protein constructs were surveyed in cell lines, and 16 constructs then were selected to generate mouse li...
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| Veröffentlicht in: | Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2011-07, Vol.49 (7), p.579-590 |
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| container_title | Genesis (New York, N.Y. : 2000) |
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| creator | Abe, Takaya Kiyonari, Hiroshi Shioi, Go Inoue, Ken-Ichi Nakao, Kazuki Aizawa, Shinichi Fujimori, Toshihiko |
| description | A series of conditional reporter mouse lines were established in which specific organelles were labeled with fluorescent proteins. Subcellular localization and intensity of 28 fluorescent fusion‐protein constructs were surveyed in cell lines, and 16 constructs then were selected to generate mouse lines. The fusion cDNAs were inserted into the ROSA26 genomic locus next to the stop sequences flanked with loxP so that fluorescent proteins were expressed under the ubiquitous ROSA26 transcriptional machinery when the loxP sequences were recombined with Cre. The subcellular localization and intensity of the fusion product in each reporter mouse line were examined by ubiquitously expressing them in E7.5 embryos. Twelve reporter lines, that mark nucleus, mitochondria, Golgi apparatus, plasma membrane, microtubule, actin filament, and focal adhesion, were found suitable for live imaging. Distinct double staining was demonstrated for nucleus and plasma membrane or Golgi apparatus; clear time‐lapse live images were obtained for nucleus and plasma membranes; conditional expression was confirmed on Lyn‐Venus and H2B‐mCherry lines in notochord with Not‐Cre. genesis, 49:579–590, 2011. © 2011 Wiley‐Liss, Inc. |
| doi_str_mv | 10.1002/dvg.20753 |
| format | Article |
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Subcellular localization and intensity of 28 fluorescent fusion‐protein constructs were surveyed in cell lines, and 16 constructs then were selected to generate mouse lines. The fusion cDNAs were inserted into the ROSA26 genomic locus next to the stop sequences flanked with loxP so that fluorescent proteins were expressed under the ubiquitous ROSA26 transcriptional machinery when the loxP sequences were recombined with Cre. The subcellular localization and intensity of the fusion product in each reporter mouse line were examined by ubiquitously expressing them in E7.5 embryos. Twelve reporter lines, that mark nucleus, mitochondria, Golgi apparatus, plasma membrane, microtubule, actin filament, and focal adhesion, were found suitable for live imaging. Distinct double staining was demonstrated for nucleus and plasma membrane or Golgi apparatus; clear time‐lapse live images were obtained for nucleus and plasma membranes; conditional expression was confirmed on Lyn‐Venus and H2B‐mCherry lines in notochord with Not‐Cre. genesis, 49:579–590, 2011. © 2011 Wiley‐Liss, Inc.</description><identifier>ISSN: 1526-954X</identifier><identifier>ISSN: 1526-968X</identifier><identifier>EISSN: 1526-968X</identifier><identifier>DOI: 10.1002/dvg.20753</identifier><identifier>PMID: 21445964</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Actin ; Animals ; Cell Line ; cell organelle ; Computed tomography ; Cre/loxP ; Dogs ; Embryo, Mammalian - cytology ; Embryo, Mammalian - metabolism ; Embryos ; Filaments ; Gene Expression Regulation, Developmental ; Gene Order ; Gene Targeting ; Genes, Reporter ; Genetic Loci - genetics ; Genetic Vectors - genetics ; genomics ; Golgi apparatus ; Intracellular Space ; live-imaging ; Luminescent Proteins - genetics ; Luminescent Proteins - metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microtubules ; Mitochondria ; Molecular Imaging ; NIH 3T3 Cells ; Notochord ; Nuclei ; Organ Specificity - genetics ; Organelles ; Plasma membranes ; Protein Transport - genetics ; Proteins - genetics ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; reporter mice ; RNA, Untranslated ; ROSA26 ; Staining and Labeling ; Transcription</subject><ispartof>Genesis (New York, N.Y. : 2000), 2011-07, Vol.49 (7), p.579-590</ispartof><rights>Copyright © 2011 Wiley‐Liss, Inc.</rights><rights>Copyright © 2011 Wiley-Liss, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4963-354a0323b621b71ab8af20ed2d53d573bbbf041c3842fcafda7f93ddbde1a3f23</citedby><cites>FETCH-LOGICAL-c4963-354a0323b621b71ab8af20ed2d53d573bbbf041c3842fcafda7f93ddbde1a3f23</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%2Fdvg.20753$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fdvg.20753$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1416,27922,27923,45572,45573</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21445964$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Abe, Takaya</creatorcontrib><creatorcontrib>Kiyonari, Hiroshi</creatorcontrib><creatorcontrib>Shioi, Go</creatorcontrib><creatorcontrib>Inoue, Ken-Ichi</creatorcontrib><creatorcontrib>Nakao, Kazuki</creatorcontrib><creatorcontrib>Aizawa, Shinichi</creatorcontrib><creatorcontrib>Fujimori, Toshihiko</creatorcontrib><title>Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging</title><title>Genesis (New York, N.Y. : 2000)</title><addtitle>Genesis</addtitle><description>A series of conditional reporter mouse lines were established in which specific organelles were labeled with fluorescent proteins. Subcellular localization and intensity of 28 fluorescent fusion‐protein constructs were surveyed in cell lines, and 16 constructs then were selected to generate mouse lines. The fusion cDNAs were inserted into the ROSA26 genomic locus next to the stop sequences flanked with loxP so that fluorescent proteins were expressed under the ubiquitous ROSA26 transcriptional machinery when the loxP sequences were recombined with Cre. The subcellular localization and intensity of the fusion product in each reporter mouse line were examined by ubiquitously expressing them in E7.5 embryos. Twelve reporter lines, that mark nucleus, mitochondria, Golgi apparatus, plasma membrane, microtubule, actin filament, and focal adhesion, were found suitable for live imaging. Distinct double staining was demonstrated for nucleus and plasma membrane or Golgi apparatus; clear time‐lapse live images were obtained for nucleus and plasma membranes; conditional expression was confirmed on Lyn‐Venus and H2B‐mCherry lines in notochord with Not‐Cre. genesis, 49:579–590, 2011. © 2011 Wiley‐Liss, Inc.</description><subject>Actin</subject><subject>Animals</subject><subject>Cell Line</subject><subject>cell organelle</subject><subject>Computed tomography</subject><subject>Cre/loxP</subject><subject>Dogs</subject><subject>Embryo, Mammalian - cytology</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryos</subject><subject>Filaments</subject><subject>Gene Expression Regulation, Developmental</subject><subject>Gene Order</subject><subject>Gene Targeting</subject><subject>Genes, Reporter</subject><subject>Genetic Loci - genetics</subject><subject>Genetic Vectors - genetics</subject><subject>genomics</subject><subject>Golgi apparatus</subject><subject>Intracellular Space</subject><subject>live-imaging</subject><subject>Luminescent Proteins - genetics</subject><subject>Luminescent Proteins - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Transgenic</subject><subject>Microtubules</subject><subject>Mitochondria</subject><subject>Molecular Imaging</subject><subject>NIH 3T3 Cells</subject><subject>Notochord</subject><subject>Nuclei</subject><subject>Organ Specificity - genetics</subject><subject>Organelles</subject><subject>Plasma membranes</subject><subject>Protein Transport - genetics</subject><subject>Proteins - genetics</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>reporter mice</subject><subject>RNA, Untranslated</subject><subject>ROSA26</subject><subject>Staining and Labeling</subject><subject>Transcription</subject><issn>1526-954X</issn><issn>1526-968X</issn><issn>1526-968X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp90MtOGzEUBmCroio0dNEXqLyDLgZ8GdszS8QlrRQlXJvsLHtsB8PMONgTWt6-E0Kyg5Ut-Tu_jn8AvmN0hBEix-Z5fkSQYPQT2MOM8KzkxWxnc2f5bBd8TekBIcQKQr6AXYLznJU83wPT89QpXft039i2g8HBKrTGdz60qobRLkLsbIRNWCYLa9_aBFUHryc3J4TDOlTLBF2I_cuzhZWta-gbNfftfB98dqpO9tvbOQB3F-e3p7-y0WT4-_RklFV5yWlGWa4QJVRzgrXAShfKEWQNMYwaJqjW2qEcV7TIiauUM0q4khqjjcWKOkIH4GCdu4jhaWlTJxufVouo1vY7y0IwQQjCRS8PP5QYYVEKhviK_lzTKoaUonVyEft_xZceyVXjsm9cvjbe2x9vsUvdWLOVm4p7cLwGf31tX95Pkmd_hpvIbD3hU2f_bSdUfJRcUMHkdDyUY359NuOXV3JG_wOXC5oH</recordid><startdate>201107</startdate><enddate>201107</enddate><creator>Abe, Takaya</creator><creator>Kiyonari, Hiroshi</creator><creator>Shioi, Go</creator><creator>Inoue, Ken-Ichi</creator><creator>Nakao, Kazuki</creator><creator>Aizawa, Shinichi</creator><creator>Fujimori, Toshihiko</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>201107</creationdate><title>Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging</title><author>Abe, Takaya ; Kiyonari, Hiroshi ; Shioi, Go ; Inoue, Ken-Ichi ; Nakao, Kazuki ; Aizawa, Shinichi ; Fujimori, Toshihiko</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4963-354a0323b621b71ab8af20ed2d53d573bbbf041c3842fcafda7f93ddbde1a3f23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Actin</topic><topic>Animals</topic><topic>Cell Line</topic><topic>cell organelle</topic><topic>Computed tomography</topic><topic>Cre/loxP</topic><topic>Dogs</topic><topic>Embryo, Mammalian - cytology</topic><topic>Embryo, Mammalian - metabolism</topic><topic>Embryos</topic><topic>Filaments</topic><topic>Gene Expression Regulation, Developmental</topic><topic>Gene Order</topic><topic>Gene Targeting</topic><topic>Genes, Reporter</topic><topic>Genetic Loci - genetics</topic><topic>Genetic Vectors - genetics</topic><topic>genomics</topic><topic>Golgi apparatus</topic><topic>Intracellular Space</topic><topic>live-imaging</topic><topic>Luminescent Proteins - genetics</topic><topic>Luminescent Proteins - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Transgenic</topic><topic>Microtubules</topic><topic>Mitochondria</topic><topic>Molecular Imaging</topic><topic>NIH 3T3 Cells</topic><topic>Notochord</topic><topic>Nuclei</topic><topic>Organ Specificity - genetics</topic><topic>Organelles</topic><topic>Plasma membranes</topic><topic>Protein Transport - genetics</topic><topic>Proteins - genetics</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>reporter mice</topic><topic>RNA, Untranslated</topic><topic>ROSA26</topic><topic>Staining and Labeling</topic><topic>Transcription</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Abe, Takaya</creatorcontrib><creatorcontrib>Kiyonari, Hiroshi</creatorcontrib><creatorcontrib>Shioi, Go</creatorcontrib><creatorcontrib>Inoue, Ken-Ichi</creatorcontrib><creatorcontrib>Nakao, Kazuki</creatorcontrib><creatorcontrib>Aizawa, Shinichi</creatorcontrib><creatorcontrib>Fujimori, Toshihiko</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>Abe, Takaya</au><au>Kiyonari, Hiroshi</au><au>Shioi, Go</au><au>Inoue, Ken-Ichi</au><au>Nakao, Kazuki</au><au>Aizawa, Shinichi</au><au>Fujimori, Toshihiko</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging</atitle><jtitle>Genesis (New York, N.Y. : 2000)</jtitle><addtitle>Genesis</addtitle><date>2011-07</date><risdate>2011</risdate><volume>49</volume><issue>7</issue><spage>579</spage><epage>590</epage><pages>579-590</pages><issn>1526-954X</issn><issn>1526-968X</issn><eissn>1526-968X</eissn><abstract>A series of conditional reporter mouse lines were established in which specific organelles were labeled with fluorescent proteins. Subcellular localization and intensity of 28 fluorescent fusion‐protein constructs were surveyed in cell lines, and 16 constructs then were selected to generate mouse lines. The fusion cDNAs were inserted into the ROSA26 genomic locus next to the stop sequences flanked with loxP so that fluorescent proteins were expressed under the ubiquitous ROSA26 transcriptional machinery when the loxP sequences were recombined with Cre. The subcellular localization and intensity of the fusion product in each reporter mouse line were examined by ubiquitously expressing them in E7.5 embryos. Twelve reporter lines, that mark nucleus, mitochondria, Golgi apparatus, plasma membrane, microtubule, actin filament, and focal adhesion, were found suitable for live imaging. Distinct double staining was demonstrated for nucleus and plasma membrane or Golgi apparatus; clear time‐lapse live images were obtained for nucleus and plasma membranes; conditional expression was confirmed on Lyn‐Venus and H2B‐mCherry lines in notochord with Not‐Cre. genesis, 49:579–590, 2011. © 2011 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>21445964</pmid><doi>10.1002/dvg.20753</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Genesis (New York, N.Y. : 2000), 2011-07, Vol.49 (7), p.579-590 |
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| subjects | Actin Animals Cell Line cell organelle Computed tomography Cre/loxP Dogs Embryo, Mammalian - cytology Embryo, Mammalian - metabolism Embryos Filaments Gene Expression Regulation, Developmental Gene Order Gene Targeting Genes, Reporter Genetic Loci - genetics Genetic Vectors - genetics genomics Golgi apparatus Intracellular Space live-imaging Luminescent Proteins - genetics Luminescent Proteins - metabolism Mice Mice, Inbred C57BL Mice, Transgenic Microtubules Mitochondria Molecular Imaging NIH 3T3 Cells Notochord Nuclei Organ Specificity - genetics Organelles Plasma membranes Protein Transport - genetics Proteins - genetics Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism reporter mice RNA, Untranslated ROSA26 Staining and Labeling Transcription |
| title | Establishment of conditional reporter mouse lines at ROSA26 locus for live cell imaging |
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