Efficient transgenesis in farm animals by lentiviral vectors
Microinjection of DNA is now the most widespread method for generating transgenic animals, but transgenesis rates achieved this way in higher mammals are extremely low. To address this longstanding problem, we used lentiviral vectors carrying a ubiquitously active promoter (phosphoglycerate kinase,...
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| Veröffentlicht in: | EMBO reports 2003-11, Vol.4 (11), p.1054-1058 |
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| creator | Hofmann, Andreas Kessler, Barbara Ewerling, Sonja Weppert, Myriam Vogg, Barbara Ludwig, Harald Stojkovic, Miodrag Boelhauve, Marc Brem, Gottfried Wolf, Eckhard Pfeifer, Alexander |
| description | Microinjection of DNA is now the most widespread method for generating transgenic animals, but transgenesis rates achieved this way in higher mammals are extremely low. To address this longstanding problem, we used lentiviral vectors carrying a ubiquitously active promoter (phosphoglycerate kinase, LV‐PGK) to deliver transgenes to porcine embryos. Of the 46 piglets born, 32 (70%) carried the transgene DNA and 30 (94%) of these pigs expressed the transgene (green fluorescent protein, GFP). Direct fluorescence imaging and immunohistochemistry showed that GFP was expressed in all tissues of LV‐PGK transgenic pigs, including germ cells. Importantly, the transgene was transmitted through the germ‐line. Tissue‐specific transgene expression was achieved by infecting porcine embryos with lentiviral vectors containing the human keratin K14 promoter (LV‐K14). LV‐K14 transgenic animals expressed GFP specifically in basal keratinocytes of the skin. Finally, infection of bovine oocytes after and before
in vitro
fertilization with LV‐PGK resulted in transgene expression in 45% and 92% of the infected embryos, respectively. |
| doi_str_mv | 10.1038/sj.embor.7400007 |
| format | Article |
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in vitro
fertilization with LV‐PGK resulted in transgene expression in 45% and 92% of the infected embryos, respectively.</description><identifier>ISSN: 1469-221X</identifier><identifier>EISSN: 1469-3178</identifier><identifier>EISSN: 1469-221X</identifier><identifier>DOI: 10.1038/sj.embor.7400007</identifier><identifier>PMID: 14566324</identifier><identifier>CODEN: ERMEAX</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Animals ; Animals, Genetically Modified ; Cattle ; Deoxyribonucleic acid ; DNA ; Embryo, Mammalian - metabolism ; Embryos ; Expression vectors ; Farms ; Fertilization ; Fluorescence ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Germ cells ; Green fluorescent protein ; imaging ; Immunohistochemistry ; In vitro fertilization ; Infection ; Keratin ; Keratinocytes ; Lentivirus ; Mammals ; Microinjection ; Microscopy, Fluorescence ; Oocytes ; Oocytes - metabolism ; Phosphoglycerate kinase ; Promoters ; Scientific Report ; Skin ; Swine ; Swine - genetics ; Transgenes ; Transgenic animals</subject><ispartof>EMBO reports, 2003-11, Vol.4 (11), p.1054-1058</ispartof><rights>European Molecular Biology Organization 2003</rights><rights>Copyright © 2003 European Molecular Biology Organization</rights><rights>Copyright Nature Publishing Group Oct 2003</rights><rights>Copyright © 2003, European Molecular Biology Organization 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6787-c0298f51de3c1a7966634bf2b8a4613b82ec0904a759cd0be204399796d309df3</citedby><cites>FETCH-LOGICAL-c6787-c0298f51de3c1a7966634bf2b8a4613b82ec0904a759cd0be204399796d309df3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1326377/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1326377/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,1411,1427,27901,27902,45550,45551,46384,46808,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14566324$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hofmann, Andreas</creatorcontrib><creatorcontrib>Kessler, Barbara</creatorcontrib><creatorcontrib>Ewerling, Sonja</creatorcontrib><creatorcontrib>Weppert, Myriam</creatorcontrib><creatorcontrib>Vogg, Barbara</creatorcontrib><creatorcontrib>Ludwig, Harald</creatorcontrib><creatorcontrib>Stojkovic, Miodrag</creatorcontrib><creatorcontrib>Boelhauve, Marc</creatorcontrib><creatorcontrib>Brem, Gottfried</creatorcontrib><creatorcontrib>Wolf, Eckhard</creatorcontrib><creatorcontrib>Pfeifer, Alexander</creatorcontrib><title>Efficient transgenesis in farm animals by lentiviral vectors</title><title>EMBO reports</title><addtitle>EMBO Rep</addtitle><addtitle>EMBO Rep</addtitle><description>Microinjection of DNA is now the most widespread method for generating transgenic animals, but transgenesis rates achieved this way in higher mammals are extremely low. To address this longstanding problem, we used lentiviral vectors carrying a ubiquitously active promoter (phosphoglycerate kinase, LV‐PGK) to deliver transgenes to porcine embryos. Of the 46 piglets born, 32 (70%) carried the transgene DNA and 30 (94%) of these pigs expressed the transgene (green fluorescent protein, GFP). Direct fluorescence imaging and immunohistochemistry showed that GFP was expressed in all tissues of LV‐PGK transgenic pigs, including germ cells. Importantly, the transgene was transmitted through the germ‐line. Tissue‐specific transgene expression was achieved by infecting porcine embryos with lentiviral vectors containing the human keratin K14 promoter (LV‐K14). LV‐K14 transgenic animals expressed GFP specifically in basal keratinocytes of the skin. Finally, infection of bovine oocytes after and before
in vitro
fertilization with LV‐PGK resulted in transgene expression in 45% and 92% of the infected embryos, respectively.</description><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Cattle</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryos</subject><subject>Expression vectors</subject><subject>Farms</subject><subject>Fertilization</subject><subject>Fluorescence</subject><subject>Gene Transfer Techniques</subject><subject>Genes, Reporter</subject><subject>Genetic Vectors</subject><subject>Germ cells</subject><subject>Green fluorescent protein</subject><subject>imaging</subject><subject>Immunohistochemistry</subject><subject>In vitro fertilization</subject><subject>Infection</subject><subject>Keratin</subject><subject>Keratinocytes</subject><subject>Lentivirus</subject><subject>Mammals</subject><subject>Microinjection</subject><subject>Microscopy, Fluorescence</subject><subject>Oocytes</subject><subject>Oocytes - metabolism</subject><subject>Phosphoglycerate kinase</subject><subject>Promoters</subject><subject>Scientific Report</subject><subject>Skin</subject><subject>Swine</subject><subject>Swine - genetics</subject><subject>Transgenes</subject><subject>Transgenic animals</subject><issn>1469-221X</issn><issn>1469-3178</issn><issn>1469-221X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkUtv1DAUhSMEoqWwZ4UiFmWVqd8PCSHR0bRQtUVCvHaW4ziDhzxaOzMw_55bEnWAReuNLd3vnHt8b5Y9x2iGEVVHaTXzbdnHmWQIjnyQ7WMmdEGxVA-nNyH42172JKUVEFxL9Tjbw4wLQQnbz14v6jq44LshH6Lt0tJ3PoWUhy6vbWxz24XWNikvt3kDUNiEaJt8493Qx_Q0e1RD0T-b7oPs88ni0_xdcf7h9P387XnhhFSycIhoVXNceeqwlVpAb1bWpFSWCUxLRbxDGjEruXYVKj1BjGoNYEWRrmp6kL0Zfa_WZesrB0EghbmKkC1uTW-D-bfShe9m2W8MpkRQKcHg1WQQ--u1T4NpQ3K-aWzn-3UymjMuIQEB8vBOUmIKwxX6XpAgwijDN44v_wNX_Tp2MC9gFMeaKwoQGiEX-5Sir28_h5G5WbVJK_Nn1WZaNUhe_D2UnWDaLQB6BH6Gxm_vNTSLi-OPO3M8ahPIuqWPu9B3BCpGTUiD_3Xbz8YfRkgqufl6eWrU5dnxXF-cmC_0N1x612Q</recordid><startdate>200311</startdate><enddate>200311</enddate><creator>Hofmann, Andreas</creator><creator>Kessler, Barbara</creator><creator>Ewerling, Sonja</creator><creator>Weppert, Myriam</creator><creator>Vogg, Barbara</creator><creator>Ludwig, Harald</creator><creator>Stojkovic, Miodrag</creator><creator>Boelhauve, Marc</creator><creator>Brem, Gottfried</creator><creator>Wolf, Eckhard</creator><creator>Pfeifer, Alexander</creator><general>John Wiley & Sons, Ltd</general><general>Nature Publishing Group UK</general><general>Springer Nature B.V</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>3V.</scope><scope>7QL</scope><scope>7T5</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>200311</creationdate><title>Efficient transgenesis in farm animals by lentiviral vectors</title><author>Hofmann, Andreas ; Kessler, Barbara ; Ewerling, Sonja ; Weppert, Myriam ; Vogg, Barbara ; Ludwig, Harald ; Stojkovic, Miodrag ; Boelhauve, Marc ; Brem, Gottfried ; Wolf, Eckhard ; Pfeifer, Alexander</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c6787-c0298f51de3c1a7966634bf2b8a4613b82ec0904a759cd0be204399796d309df3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Cattle</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>Embryo, Mammalian - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>EMBO reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hofmann, Andreas</au><au>Kessler, Barbara</au><au>Ewerling, Sonja</au><au>Weppert, Myriam</au><au>Vogg, Barbara</au><au>Ludwig, Harald</au><au>Stojkovic, Miodrag</au><au>Boelhauve, Marc</au><au>Brem, Gottfried</au><au>Wolf, Eckhard</au><au>Pfeifer, Alexander</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient transgenesis in farm animals by lentiviral vectors</atitle><jtitle>EMBO reports</jtitle><stitle>EMBO Rep</stitle><addtitle>EMBO Rep</addtitle><date>2003-11</date><risdate>2003</risdate><volume>4</volume><issue>11</issue><spage>1054</spage><epage>1058</epage><pages>1054-1058</pages><issn>1469-221X</issn><eissn>1469-3178</eissn><eissn>1469-221X</eissn><coden>ERMEAX</coden><abstract>Microinjection of DNA is now the most widespread method for generating transgenic animals, but transgenesis rates achieved this way in higher mammals are extremely low. To address this longstanding problem, we used lentiviral vectors carrying a ubiquitously active promoter (phosphoglycerate kinase, LV‐PGK) to deliver transgenes to porcine embryos. Of the 46 piglets born, 32 (70%) carried the transgene DNA and 30 (94%) of these pigs expressed the transgene (green fluorescent protein, GFP). Direct fluorescence imaging and immunohistochemistry showed that GFP was expressed in all tissues of LV‐PGK transgenic pigs, including germ cells. Importantly, the transgene was transmitted through the germ‐line. Tissue‐specific transgene expression was achieved by infecting porcine embryos with lentiviral vectors containing the human keratin K14 promoter (LV‐K14). LV‐K14 transgenic animals expressed GFP specifically in basal keratinocytes of the skin. Finally, infection of bovine oocytes after and before
in vitro
fertilization with LV‐PGK resulted in transgene expression in 45% and 92% of the infected embryos, respectively.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>14566324</pmid><doi>10.1038/sj.embor.7400007</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete; Wiley Online Library Free Content; EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Animals Animals, Genetically Modified Cattle Deoxyribonucleic acid DNA Embryo, Mammalian - metabolism Embryos Expression vectors Farms Fertilization Fluorescence Gene Transfer Techniques Genes, Reporter Genetic Vectors Germ cells Green fluorescent protein imaging Immunohistochemistry In vitro fertilization Infection Keratin Keratinocytes Lentivirus Mammals Microinjection Microscopy, Fluorescence Oocytes Oocytes - metabolism Phosphoglycerate kinase Promoters Scientific Report Skin Swine Swine - genetics Transgenes Transgenic animals |
| title | Efficient transgenesis in farm animals by lentiviral vectors |
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