Rosa26 locus supports tissue-specific promoter driving transgene expression specifically in pig
Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified...
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| creator | Kong, Qingran Hai, Tang Ma, Jing Huang, Tianqing Jiang, Dandan Xie, Bingteng Wu, Meiling Wang, Jiaqiang Song, Yuran Wang, Ying He, Yilong Sun, Jialu Hu, Kui Guo, Runfa Wang, Liu Zhou, Qi Mu, Yanshuang Liu, Zhonghua |
| description | Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 (pRosa26) locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter (pR26) was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter (EF1a) -driven GFP reporter and Myostatin promoter (MyoP)-driven Follistatin (Fst) were successfully targeted into the pRosa26 locusby traditional homologous recombination (HR) strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus. |
| doi_str_mv | 10.1371/journal.pone.0107945 |
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However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 (pRosa26) locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter (pR26) was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter (EF1a) -driven GFP reporter and Myostatin promoter (MyoP)-driven Follistatin (Fst) were successfully targeted into the pRosa26 locusby traditional homologous recombination (HR) strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0107945</identifier><identifier>PMID: 25232950</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Agricultural research ; Agriculture ; Animal genetic engineering ; Animals ; Animals, Genetically Modified ; Base Sequence ; Biology ; Biology and Life Sciences ; Biomedical materials ; Biotechnology ; Cells, Cultured ; CRISPR ; Deoxyribonucleic acid ; DNA ; DNA methylation ; Elongation ; Follistatin ; Follistatin gene ; Gene Expression ; Gene targeting ; Gene therapy ; Genes, Reporter ; Genetic Loci ; Genetic modification ; Genetically modified animals ; Green Fluorescent Proteins - biosynthesis ; Green Fluorescent Proteins - genetics ; Hogs ; Homologous recombination ; Homology ; Laboratories ; Life sciences ; Loci ; Methylation ; Molecular Sequence Data ; Muscle, Skeletal - metabolism ; Muscles ; Mutation ; Myostatin ; Neurosciences ; Organ Specificity ; Phenotypes ; Promoter Regions, Genetic ; Reporter gene ; RNA, Untranslated - genetics ; Rodents ; Stem cells ; Suidae ; Sus scrofa - genetics ; Swine ; Swine, Miniature ; Transcriptional Activation ; Transgenes ; Zoology</subject><ispartof>PloS one, 2014-09, Vol.9 (9), p.e107945-e107945</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Kong et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Kong et al 2014 Kong et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-f9e9e1ae5a8ca44e6d8bed1025b116a5805f3f9e7e0b7f70314e122ae0a198f33</citedby><cites>FETCH-LOGICAL-c758t-f9e9e1ae5a8ca44e6d8bed1025b116a5805f3f9e7e0b7f70314e122ae0a198f33</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/PMC4169413/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4169413/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,865,886,2103,2929,23868,27926,27927,53793,53795</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25232950$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>James, Leighton R.</contributor><creatorcontrib>Kong, Qingran</creatorcontrib><creatorcontrib>Hai, Tang</creatorcontrib><creatorcontrib>Ma, Jing</creatorcontrib><creatorcontrib>Huang, Tianqing</creatorcontrib><creatorcontrib>Jiang, Dandan</creatorcontrib><creatorcontrib>Xie, Bingteng</creatorcontrib><creatorcontrib>Wu, Meiling</creatorcontrib><creatorcontrib>Wang, Jiaqiang</creatorcontrib><creatorcontrib>Song, Yuran</creatorcontrib><creatorcontrib>Wang, Ying</creatorcontrib><creatorcontrib>He, Yilong</creatorcontrib><creatorcontrib>Sun, Jialu</creatorcontrib><creatorcontrib>Hu, Kui</creatorcontrib><creatorcontrib>Guo, Runfa</creatorcontrib><creatorcontrib>Wang, Liu</creatorcontrib><creatorcontrib>Zhou, Qi</creatorcontrib><creatorcontrib>Mu, Yanshuang</creatorcontrib><creatorcontrib>Liu, Zhonghua</creatorcontrib><title>Rosa26 locus supports tissue-specific promoter driving transgene expression specifically in pig</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 (pRosa26) locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter (pR26) was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter (EF1a) -driven GFP reporter and Myostatin promoter (MyoP)-driven Follistatin (Fst) were successfully targeted into the pRosa26 locusby traditional homologous recombination (HR) strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus.</description><subject>Agricultural research</subject><subject>Agriculture</subject><subject>Animal genetic engineering</subject><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Base Sequence</subject><subject>Biology</subject><subject>Biology and Life Sciences</subject><subject>Biomedical materials</subject><subject>Biotechnology</subject><subject>Cells, Cultured</subject><subject>CRISPR</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA methylation</subject><subject>Elongation</subject><subject>Follistatin</subject><subject>Follistatin gene</subject><subject>Gene Expression</subject><subject>Gene targeting</subject><subject>Gene therapy</subject><subject>Genes, Reporter</subject><subject>Genetic Loci</subject><subject>Genetic modification</subject><subject>Genetically modified animals</subject><subject>Green Fluorescent Proteins - biosynthesis</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Hogs</subject><subject>Homologous recombination</subject><subject>Homology</subject><subject>Laboratories</subject><subject>Life sciences</subject><subject>Loci</subject><subject>Methylation</subject><subject>Molecular Sequence Data</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Muscles</subject><subject>Mutation</subject><subject>Myostatin</subject><subject>Neurosciences</subject><subject>Organ Specificity</subject><subject>Phenotypes</subject><subject>Promoter Regions, Genetic</subject><subject>Reporter gene</subject><subject>RNA, Untranslated - genetics</subject><subject>Rodents</subject><subject>Stem cells</subject><subject>Suidae</subject><subject>Sus scrofa - genetics</subject><subject>Swine</subject><subject>Swine, Miniature</subject><subject>Transcriptional 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locus supports tissue-specific promoter driving transgene expression specifically in pig</title><author>Kong, Qingran ; Hai, Tang ; Ma, Jing ; Huang, Tianqing ; Jiang, Dandan ; Xie, Bingteng ; Wu, Meiling ; Wang, Jiaqiang ; Song, Yuran ; Wang, Ying ; He, Yilong ; Sun, Jialu ; Hu, Kui ; Guo, Runfa ; Wang, Liu ; Zhou, Qi ; Mu, Yanshuang ; Liu, Zhonghua</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-f9e9e1ae5a8ca44e6d8bed1025b116a5805f3f9e7e0b7f70314e122ae0a198f33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Agricultural research</topic><topic>Agriculture</topic><topic>Animal genetic engineering</topic><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Base Sequence</topic><topic>Biology</topic><topic>Biology and Life Sciences</topic><topic>Biomedical materials</topic><topic>Biotechnology</topic><topic>Cells, Cultured</topic><topic>CRISPR</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA methylation</topic><topic>Elongation</topic><topic>Follistatin</topic><topic>Follistatin gene</topic><topic>Gene Expression</topic><topic>Gene targeting</topic><topic>Gene therapy</topic><topic>Genes, Reporter</topic><topic>Genetic Loci</topic><topic>Genetic modification</topic><topic>Genetically modified animals</topic><topic>Green Fluorescent Proteins - biosynthesis</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Hogs</topic><topic>Homologous recombination</topic><topic>Homology</topic><topic>Laboratories</topic><topic>Life sciences</topic><topic>Loci</topic><topic>Methylation</topic><topic>Molecular Sequence Data</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Muscles</topic><topic>Mutation</topic><topic>Myostatin</topic><topic>Neurosciences</topic><topic>Organ Specificity</topic><topic>Phenotypes</topic><topic>Promoter Regions, 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Tang</au><au>Ma, Jing</au><au>Huang, Tianqing</au><au>Jiang, Dandan</au><au>Xie, Bingteng</au><au>Wu, Meiling</au><au>Wang, Jiaqiang</au><au>Song, Yuran</au><au>Wang, Ying</au><au>He, Yilong</au><au>Sun, Jialu</au><au>Hu, Kui</au><au>Guo, Runfa</au><au>Wang, Liu</au><au>Zhou, Qi</au><au>Mu, Yanshuang</au><au>Liu, Zhonghua</au><au>James, Leighton R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rosa26 locus supports tissue-specific promoter driving transgene expression specifically in pig</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-09-18</date><risdate>2014</risdate><volume>9</volume><issue>9</issue><spage>e107945</spage><epage>e107945</epage><pages>e107945-e107945</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 (pRosa26) locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter (pR26) was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter (EF1a) -driven GFP reporter and Myostatin promoter (MyoP)-driven Follistatin (Fst) were successfully targeted into the pRosa26 locusby traditional homologous recombination (HR) strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25232950</pmid><doi>10.1371/journal.pone.0107945</doi><oa>free_for_read</oa></addata></record> |
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| language | eng |
| recordid | cdi_plos_journals_1563048939 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Agricultural research Agriculture Animal genetic engineering Animals Animals, Genetically Modified Base Sequence Biology Biology and Life Sciences Biomedical materials Biotechnology Cells, Cultured CRISPR Deoxyribonucleic acid DNA DNA methylation Elongation Follistatin Follistatin gene Gene Expression Gene targeting Gene therapy Genes, Reporter Genetic Loci Genetic modification Genetically modified animals Green Fluorescent Proteins - biosynthesis Green Fluorescent Proteins - genetics Hogs Homologous recombination Homology Laboratories Life sciences Loci Methylation Molecular Sequence Data Muscle, Skeletal - metabolism Muscles Mutation Myostatin Neurosciences Organ Specificity Phenotypes Promoter Regions, Genetic Reporter gene RNA, Untranslated - genetics Rodents Stem cells Suidae Sus scrofa - genetics Swine Swine, Miniature Transcriptional Activation Transgenes Zoology |
| title | Rosa26 locus supports tissue-specific promoter driving transgene expression specifically in pig |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-18T04%3A49%3A49IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Rosa26%20locus%20supports%20tissue-specific%20promoter%20driving%20transgene%20expression%20specifically%20in%20pig&rft.jtitle=PloS%20one&rft.au=Kong,%20Qingran&rft.date=2014-09-18&rft.volume=9&rft.issue=9&rft.spage=e107945&rft.epage=e107945&rft.pages=e107945-e107945&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0107945&rft_dat=%3Cgale_plos_%3EA418138559%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1563048939&rft_id=info:pmid/25232950&rft_galeid=A418138559&rft_doaj_id=oai_doaj_org_article_bccb463d6dfd4412b2071b580074283b&rfr_iscdi=true |