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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2012
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.
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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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