Pig transgenesis by Sleeping Beauty DNA transposition
Modelling of human disease in genetically engineered pigs provides unique possibilities in biomedical research and in studies of disease intervention. Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease...
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| Veröffentlicht in: | Transgenic research 2011-06, Vol.20 (3), p.533-545 |
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| creator | Jakobsen, Jannik E. Li, Juan Kragh, Peter M. Moldt, Brian Lin, Lin Liu, Ying Schmidt, Mette Winther, Kjeld Dahl Schyth, Brian Dall Holm, Ida E. Vajta, Gábor Bolund, Lars Callesen, Henrik Jørgensen, Arne Lund Nielsen, Anders Lade Mikkelsen, Jacob Giehm |
| description | Modelling of human disease in genetically engineered pigs provides unique possibilities in biomedical research and in studies of disease intervention. Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology. |
| doi_str_mv | 10.1007/s11248-010-9438-x |
| format | Article |
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Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology.</description><identifier>ISSN: 0962-8819</identifier><identifier>EISSN: 1573-9368</identifier><identifier>DOI: 10.1007/s11248-010-9438-x</identifier><identifier>PMID: 20803249</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Animal Genetics and Genomics ; Animals ; Animals, Genetically Modified ; Base Sequence ; Biological and medical sciences ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biotechnology ; DNA Transposable Elements - genetics ; Fibroblasts - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene Transfer Techniques ; Genetic Engineering ; Genetic Engineering - methods ; Genetic technics ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; Humans ; Life Sciences ; Methods. Procedures. Technologies ; Molecular Medicine ; Molecular Sequence Data ; Mutagenesis, Insertional ; Original Paper ; Plant Genetics and Genomics ; Swine ; Swine, Miniature ; Transgenes - genetics ; Transgenes - physiology ; Transgenic animals and transgenic plants ; Transgenics ; Transposases - genetics ; Transposases - metabolism</subject><ispartof>Transgenic research, 2011-06, Vol.20 (3), p.533-545</ispartof><rights>Springer Science+Business Media B.V. 2010</rights><rights>2015 INIST-CNRS</rights><rights>Springer Science+Business Media B.V. 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-f9a67b1a37a84f1fd93be53e64fbf8009e667fc93c139322a417013e2c31543b3</citedby><cites>FETCH-LOGICAL-c475t-f9a67b1a37a84f1fd93be53e64fbf8009e667fc93c139322a417013e2c31543b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11248-010-9438-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11248-010-9438-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27928,27929,41492,42561,51323</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24208594$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20803249$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jakobsen, Jannik E.</creatorcontrib><creatorcontrib>Li, Juan</creatorcontrib><creatorcontrib>Kragh, Peter M.</creatorcontrib><creatorcontrib>Moldt, Brian</creatorcontrib><creatorcontrib>Lin, Lin</creatorcontrib><creatorcontrib>Liu, Ying</creatorcontrib><creatorcontrib>Schmidt, Mette</creatorcontrib><creatorcontrib>Winther, Kjeld Dahl</creatorcontrib><creatorcontrib>Schyth, Brian Dall</creatorcontrib><creatorcontrib>Holm, Ida E.</creatorcontrib><creatorcontrib>Vajta, Gábor</creatorcontrib><creatorcontrib>Bolund, Lars</creatorcontrib><creatorcontrib>Callesen, Henrik</creatorcontrib><creatorcontrib>Jørgensen, Arne Lund</creatorcontrib><creatorcontrib>Nielsen, Anders Lade</creatorcontrib><creatorcontrib>Mikkelsen, Jacob Giehm</creatorcontrib><title>Pig transgenesis by Sleeping Beauty DNA transposition</title><title>Transgenic research</title><addtitle>Transgenic Res</addtitle><addtitle>Transgenic Res</addtitle><description>Modelling of human disease in genetically engineered pigs provides unique possibilities in biomedical research and in studies of disease intervention. Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology.</description><subject>Animal Genetics and Genomics</subject><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>Biotechnology</subject><subject>DNA Transposable Elements - genetics</subject><subject>Fibroblasts - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Transfer Techniques</subject><subject>Genetic Engineering</subject><subject>Genetic Engineering - methods</subject><subject>Genetic technics</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular Medicine</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Insertional</subject><subject>Original Paper</subject><subject>Plant Genetics and Genomics</subject><subject>Swine</subject><subject>Swine, Miniature</subject><subject>Transgenes - genetics</subject><subject>Transgenes - physiology</subject><subject>Transgenic animals and transgenic plants</subject><subject>Transgenics</subject><subject>Transposases - genetics</subject><subject>Transposases - metabolism</subject><issn>0962-8819</issn><issn>1573-9368</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp10F1LwzAUBuAgipvTH-CNFEG8qubko0ku5_yEoYJ6HdIuKR1dW5sWtn9vRqeC4FUuznPeHF6ETgFfAcbi2gMQJmMMOFaMyni9h8bABY0VTeQ-GmOVkFhKUCN05P0SB4glPUQjgiWmhKkx4q9FHnWtqXxuK-sLH6Wb6K20timqPLqxpu820e3zdDBN7YuuqKtjdOBM6e3J7p2gj_u799ljPH95eJpN53HGBO9ip0wiUjBUGMkcuIWiqeXUJsylTmKsbJIIlymaAVWUEMNAYKCWZBQ4oymdoMsht2nrz976Tq8Kn9myNJWte6-lUpBIQnCQ53_ksu7bKhynpeCcYQ5bBAPK2tr71jrdtMXKtBsNWG8b1UOjOhSlt43qddg52wX36coufja-KwzgYgeMz0zpQk9Z4X8dC5KHsAkig_NhVOW2_b3w_9-_ABp0jCY</recordid><startdate>20110601</startdate><enddate>20110601</enddate><creator>Jakobsen, Jannik E.</creator><creator>Li, Juan</creator><creator>Kragh, Peter M.</creator><creator>Moldt, Brian</creator><creator>Lin, Lin</creator><creator>Liu, Ying</creator><creator>Schmidt, Mette</creator><creator>Winther, Kjeld Dahl</creator><creator>Schyth, Brian Dall</creator><creator>Holm, Ida E.</creator><creator>Vajta, Gábor</creator><creator>Bolund, Lars</creator><creator>Callesen, Henrik</creator><creator>Jørgensen, Arne Lund</creator><creator>Nielsen, Anders Lade</creator><creator>Mikkelsen, Jacob Giehm</creator><general>Springer Netherlands</general><general>Springer</general><general>Springer Nature B.V</general><scope>IQODW</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>7TK</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7QO</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>20110601</creationdate><title>Pig transgenesis by Sleeping Beauty DNA transposition</title><author>Jakobsen, Jannik E. ; Li, Juan ; Kragh, Peter M. ; Moldt, Brian ; Lin, Lin ; Liu, Ying ; Schmidt, Mette ; Winther, Kjeld Dahl ; Schyth, Brian Dall ; Holm, Ida E. ; Vajta, Gábor ; Bolund, Lars ; Callesen, Henrik ; Jørgensen, Arne Lund ; Nielsen, Anders Lade ; Mikkelsen, Jacob Giehm</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-f9a67b1a37a84f1fd93be53e64fbf8009e667fc93c139322a417013e2c31543b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animal Genetics and Genomics</topic><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedical Engineering/Biotechnology</topic><topic>Biotechnology</topic><topic>DNA Transposable Elements - genetics</topic><topic>Fibroblasts - metabolism</topic><topic>Fundamental and applied biological sciences. 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Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>20803249</pmid><doi>10.1007/s11248-010-9438-x</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Transgenic research, 2011-06, Vol.20 (3), p.533-545 |
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| subjects | Animal Genetics and Genomics Animals Animals, Genetically Modified Base Sequence Biological and medical sciences Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biotechnology DNA Transposable Elements - genetics Fibroblasts - metabolism Fundamental and applied biological sciences. Psychology Gene Transfer Techniques Genetic Engineering Genetic Engineering - methods Genetic technics Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Humans Life Sciences Methods. Procedures. Technologies Molecular Medicine Molecular Sequence Data Mutagenesis, Insertional Original Paper Plant Genetics and Genomics Swine Swine, Miniature Transgenes - genetics Transgenes - physiology Transgenic animals and transgenic plants Transgenics Transposases - genetics Transposases - metabolism |
| title | Pig transgenesis by Sleeping Beauty DNA transposition |
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