Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector
Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo‐cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSC...
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| creator | Uhm, Sang Jun Gupta, Mukesh Kumar Kim, Teoan Lee, Hoon Taek |
| description | Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo‐cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine‐ and bovine‐cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNβ‐EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P |
| doi_str_mv | 10.1002/mrd.20755 |
| format | Article |
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This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine‐ and bovine‐cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNβ‐EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P < 0.05) development rates of the intraspecies SCNT porcine or bovine embryos did not differ between transgenic and nontransgenic groups. Expression of EGFP was observed in 100% of blastocysts and mosaicism was not observed. Furthermore, after iSCNT of porcine or bovine donor nuclei into xenogenic ooplasm, patterns of nuclear remodeling in reconstructed embryos were similar. In conclusion, our data demonstrated the feasibility of producing transgenic iSCNT embryos. To our knowledge, this is the first report of transgenic cloned embryo production by iSCNT approach. In the future, this may provide a powerful research tool for studying developmental events in domestic animals and provide marked cell lines for other genetic manipulations. Mol. Reprod. Dev. 74: 1538–1547, 2007. © 2007 Wiley‐Liss, Inc.</description><identifier>ISSN: 1040-452X</identifier><identifier>EISSN: 1098-2795</identifier><identifier>DOI: 10.1002/mrd.20755</identifier><identifier>PMID: 17492765</identifier><identifier>CODEN: MREDEE</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Biological and medical sciences ; Biotechnology ; Cattle - embryology ; Cell Cycle ; Chromatin - ultrastructure ; Cloning, Organism ; EGFP ; Embryo, Mammalian - chemistry ; Embryo, Mammalian - metabolism ; Embryonic Development ; Female ; Fibroblasts - chemistry ; Fibroblasts - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetic engineering ; Genetic technics ; Genetic Vectors - genetics ; Green Fluorescent Proteins - analysis ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; iSCNT ; Methods. Procedures. Technologies ; nuclear transfer ; Nuclear Transfer Techniques ; pig ; Retroviridae ; Swine - embryology ; Transfection ; transgenesis ; Vectors (cloning, transfer, expression). Insertion sequences and transposons</subject><ispartof>Molecular reproduction and development, 2007-12, Vol.74 (12), p.1538-1547</ispartof><rights>Copyright © 2007 Wiley‐Liss, Inc.</rights><rights>2008 INIST-CNRS</rights><rights>(c) 2007 Wiley-Liss, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4885-a58487e06937782da4b71c64f3d60945ab54b58f8e65ce5ead61e2d0aa757fd33</citedby><cites>FETCH-LOGICAL-c4885-a58487e06937782da4b71c64f3d60945ab54b58f8e65ce5ead61e2d0aa757fd33</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%2Fmrd.20755$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fmrd.20755$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19172174$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17492765$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Uhm, Sang Jun</creatorcontrib><creatorcontrib>Gupta, Mukesh Kumar</creatorcontrib><creatorcontrib>Kim, Teoan</creatorcontrib><creatorcontrib>Lee, Hoon Taek</creatorcontrib><title>Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector</title><title>Molecular reproduction and development</title><addtitle>Mol. Reprod. Dev</addtitle><description>Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo‐cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine‐ and bovine‐cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNβ‐EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P < 0.05) development rates of the intraspecies SCNT porcine or bovine embryos did not differ between transgenic and nontransgenic groups. Expression of EGFP was observed in 100% of blastocysts and mosaicism was not observed. Furthermore, after iSCNT of porcine or bovine donor nuclei into xenogenic ooplasm, patterns of nuclear remodeling in reconstructed embryos were similar. In conclusion, our data demonstrated the feasibility of producing transgenic iSCNT embryos. To our knowledge, this is the first report of transgenic cloned embryo production by iSCNT approach. In the future, this may provide a powerful research tool for studying developmental events in domestic animals and provide marked cell lines for other genetic manipulations. Mol. Reprod. Dev. 74: 1538–1547, 2007. © 2007 Wiley‐Liss, Inc.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cattle - embryology</subject><subject>Cell Cycle</subject><subject>Chromatin - ultrastructure</subject><subject>Cloning, Organism</subject><subject>EGFP</subject><subject>Embryo, Mammalian - chemistry</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryonic Development</subject><subject>Female</subject><subject>Fibroblasts - chemistry</subject><subject>Fibroblasts - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Vectors - genetics</subject><subject>Green Fluorescent Proteins - analysis</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>iSCNT</subject><subject>Methods. Procedures. Technologies</subject><subject>nuclear transfer</subject><subject>Nuclear Transfer Techniques</subject><subject>pig</subject><subject>Retroviridae</subject><subject>Swine - embryology</subject><subject>Transfection</subject><subject>transgenesis</subject><subject>Vectors (cloning, transfer, expression). Insertion sequences and transposons</subject><issn>1040-452X</issn><issn>1098-2795</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkdFuFSEQhjdGY2v1whcw3GjixbawCwt7aY5tNak1NRq9Iyw7VJSFLbBtz7P5cnI8p_bKmJAwGb75Z5i_qp4TfEgwbo6mOB42mDP2oNonuBd1w3v2cBNTXFPWfNurnqT0A2Pc9wI_rvYIp33DO7Zf_Tq-nSOkZINHwSDw35XXMKLLCOCRcUsorxp8RnMMGaxH5cwhauuhRsqPaAjXm1i74EsdTENch4RMcC7cWH9Z-AwxzaAtJJTCpLLVSINzyC_agYooR-WTgbgZwNghhsGplNNdXueiO6xRhBxLr7gkdF2SIT6tHhnlEjzb3QfVl5Pjz6t39dnH0_erN2e1pkKwWjFBBQfc9S3nohkVHTjRHTXt2OGeMjUwOjBhBHRMAwM1dgSaESvFGTdj2x5Ur7a6ZQVXC6QsJ5s2P1AewpJkJ1pCupb-F2xwR2nPeAFfb0EdQ0oRjJyjnVRcS4LlxlJZLJV_LC3si53oMkww3pM7DwvwcgeopJUzZWvapnuuJ7wpcOGOttyNdbD-d0f54dPbu9b1tsKmDLd_K1T8KTvecia_np9Kcn5yQVeilxftbzs4zNk</recordid><startdate>200712</startdate><enddate>200712</enddate><creator>Uhm, Sang Jun</creator><creator>Gupta, Mukesh Kumar</creator><creator>Kim, Teoan</creator><creator>Lee, Hoon Taek</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>BSCLL</scope><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>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>200712</creationdate><title>Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector</title><author>Uhm, Sang Jun ; Gupta, Mukesh Kumar ; Kim, Teoan ; Lee, Hoon Taek</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4885-a58487e06937782da4b71c64f3d60945ab54b58f8e65ce5ead61e2d0aa757fd33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cattle - embryology</topic><topic>Cell Cycle</topic><topic>Chromatin - ultrastructure</topic><topic>Cloning, Organism</topic><topic>EGFP</topic><topic>Embryo, Mammalian - chemistry</topic><topic>Embryo, Mammalian - metabolism</topic><topic>Embryonic Development</topic><topic>Female</topic><topic>Fibroblasts - chemistry</topic><topic>Fibroblasts - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genetic Vectors - genetics</topic><topic>Green Fluorescent Proteins - analysis</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>iSCNT</topic><topic>Methods. Procedures. Technologies</topic><topic>nuclear transfer</topic><topic>Nuclear Transfer Techniques</topic><topic>pig</topic><topic>Retroviridae</topic><topic>Swine - embryology</topic><topic>Transfection</topic><topic>transgenesis</topic><topic>Vectors (cloning, transfer, expression). Insertion sequences and transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Uhm, Sang Jun</creatorcontrib><creatorcontrib>Gupta, Mukesh Kumar</creatorcontrib><creatorcontrib>Kim, Teoan</creatorcontrib><creatorcontrib>Lee, Hoon Taek</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular reproduction and development</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Uhm, Sang Jun</au><au>Gupta, Mukesh Kumar</au><au>Kim, Teoan</au><au>Lee, Hoon Taek</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector</atitle><jtitle>Molecular reproduction and development</jtitle><addtitle>Mol. Reprod. Dev</addtitle><date>2007-12</date><risdate>2007</risdate><volume>74</volume><issue>12</issue><spage>1538</spage><epage>1547</epage><pages>1538-1547</pages><issn>1040-452X</issn><eissn>1098-2795</eissn><coden>MREDEE</coden><abstract>Interspecies somatic cell nuclear transfer (iSCNT) has emerged as an important tool for studying nucleo‐cytoplasmic interactions and cloning of animals whose oocytes are difficult to obtain. This study was designed to explore the feasibility of employing transgenic fibroblasts as donor cells for iSCNT. The study examined the chromatin morphology, in vitro development, and expression of an enhanced green fluorescent protein (EGFP) gene in porcine‐ and bovine‐cloned embryos produced by iSCNT of fetal fibroblast transfected with a pLNβ‐EGFP retroviral vector. Parthenogenetic and transfected or nontransfected intraspecies SCNT embryos were used as controls for comparison. Analysis of data revealed that xenogenic oocyte was able to reprogram somatic cells of different genus and supports their in vitro development to the blastocyst stage. However, the developmental rates of transgenic iSCNT embryos to the blastocyst stage were significantly lower than those of intraspecies SCNT embryos. The reduction in development rates was however, not due to integration of the transgene as the lower (P < 0.05) development rates of the intraspecies SCNT porcine or bovine embryos did not differ between transgenic and nontransgenic groups. Expression of EGFP was observed in 100% of blastocysts and mosaicism was not observed. Furthermore, after iSCNT of porcine or bovine donor nuclei into xenogenic ooplasm, patterns of nuclear remodeling in reconstructed embryos were similar. In conclusion, our data demonstrated the feasibility of producing transgenic iSCNT embryos. To our knowledge, this is the first report of transgenic cloned embryo production by iSCNT approach. In the future, this may provide a powerful research tool for studying developmental events in domestic animals and provide marked cell lines for other genetic manipulations. Mol. Reprod. Dev. 74: 1538–1547, 2007. © 2007 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>17492765</pmid><doi>10.1002/mrd.20755</doi><tpages>10</tpages></addata></record> |
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| subjects | Animals Biological and medical sciences Biotechnology Cattle - embryology Cell Cycle Chromatin - ultrastructure Cloning, Organism EGFP Embryo, Mammalian - chemistry Embryo, Mammalian - metabolism Embryonic Development Female Fibroblasts - chemistry Fibroblasts - metabolism Fundamental and applied biological sciences. Psychology Gene Expression Genetic engineering Genetic technics Genetic Vectors - genetics Green Fluorescent Proteins - analysis Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism iSCNT Methods. Procedures. Technologies nuclear transfer Nuclear Transfer Techniques pig Retroviridae Swine - embryology Transfection transgenesis Vectors (cloning, transfer, expression). Insertion sequences and transposons |
| title | Expression of enhanced green fluorescent protein in porcine- and bovine-cloned embryos following interspecies somatic cell nuclear transfer of fibroblasts transfected by retrovirus vector |
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