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Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT)
Targeted transgenesis using site-specific recombinases is an attractive method to create genetically modified animals as it allows for integration of the transgene in a pre-selected transcriptionally active genomic site. Here we describe the application of recombinase-mediated cassette exchange (RMC...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712138/ https://www.ncbi.nlm.nih.gov/pubmed/23111619 http://dx.doi.org/10.1007/s11248-012-9671-6 |
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author | Jakobsen, Jannik Ejnar Johansen, Marianne G. Schmidt, Mette Dagnæs-Hansen, Frederik Dam, Karen Gunnarsson, Anders Liu, Ying Kragh, Peter M. Li, Rong Holm, Ida E. Callesen, Henrik Mikkelsen, Jacob Giehm Nielsen, Anders Lade Jørgensen, Arne Lund |
author_facet | Jakobsen, Jannik Ejnar Johansen, Marianne G. Schmidt, Mette Dagnæs-Hansen, Frederik Dam, Karen Gunnarsson, Anders Liu, Ying Kragh, Peter M. Li, Rong Holm, Ida E. Callesen, Henrik Mikkelsen, Jacob Giehm Nielsen, Anders Lade Jørgensen, Arne Lund |
author_sort | Jakobsen, Jannik Ejnar |
collection | PubMed |
description | Targeted transgenesis using site-specific recombinases is an attractive method to create genetically modified animals as it allows for integration of the transgene in a pre-selected transcriptionally active genomic site. Here we describe the application of recombinase-mediated cassette exchange (RMCE) in cells from a Göttingen minipig with four RMCE acceptor loci, each containing a green fluorescence protein (GFP) marker gene driven by a human UbiC promoter. The four RMCE acceptor loci segregated independent of each other, and expression profiles could be determined in various tissues. Using minicircles in RMCE in fibroblasts with all four acceptor loci and followed by SCNT, we produced piglets with a single copy of a transgene incorporated into one of the transcriptionally active acceptor loci. The transgene, consisting of a cDNA of the Alzheimer’s disease-causing gene PSEN1M146I driven by an enhanced human UbiC promoter, had an expression profile in various tissues similar to that of the GFP marker gene. The results show that RMCE can be done in a pre-selected transcriptionally active acceptor locus for targeted transgenesis in pigs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11248-012-9671-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-3712138 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-37121382013-07-18 Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) Jakobsen, Jannik Ejnar Johansen, Marianne G. Schmidt, Mette Dagnæs-Hansen, Frederik Dam, Karen Gunnarsson, Anders Liu, Ying Kragh, Peter M. Li, Rong Holm, Ida E. Callesen, Henrik Mikkelsen, Jacob Giehm Nielsen, Anders Lade Jørgensen, Arne Lund Transgenic Res Original Paper Targeted transgenesis using site-specific recombinases is an attractive method to create genetically modified animals as it allows for integration of the transgene in a pre-selected transcriptionally active genomic site. Here we describe the application of recombinase-mediated cassette exchange (RMCE) in cells from a Göttingen minipig with four RMCE acceptor loci, each containing a green fluorescence protein (GFP) marker gene driven by a human UbiC promoter. The four RMCE acceptor loci segregated independent of each other, and expression profiles could be determined in various tissues. Using minicircles in RMCE in fibroblasts with all four acceptor loci and followed by SCNT, we produced piglets with a single copy of a transgene incorporated into one of the transcriptionally active acceptor loci. The transgene, consisting of a cDNA of the Alzheimer’s disease-causing gene PSEN1M146I driven by an enhanced human UbiC promoter, had an expression profile in various tissues similar to that of the GFP marker gene. The results show that RMCE can be done in a pre-selected transcriptionally active acceptor locus for targeted transgenesis in pigs. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11248-012-9671-6) contains supplementary material, which is available to authorized users. Springer Netherlands 2012-10-31 2013 /pmc/articles/PMC3712138/ /pubmed/23111619 http://dx.doi.org/10.1007/s11248-012-9671-6 Text en © The Author(s) 2012 https://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Paper Jakobsen, Jannik Ejnar Johansen, Marianne G. Schmidt, Mette Dagnæs-Hansen, Frederik Dam, Karen Gunnarsson, Anders Liu, Ying Kragh, Peter M. Li, Rong Holm, Ida E. Callesen, Henrik Mikkelsen, Jacob Giehm Nielsen, Anders Lade Jørgensen, Arne Lund Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title | Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title_full | Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title_fullStr | Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title_full_unstemmed | Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title_short | Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT) |
title_sort | generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (rmce) and somatic cell nuclear transfer (scnt) |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712138/ https://www.ncbi.nlm.nih.gov/pubmed/23111619 http://dx.doi.org/10.1007/s11248-012-9671-6 |
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