Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT).
Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT).
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DOI：
10.1007/s11248-012-9671-6
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发表时间：
2013-08
影响因子：
3
通讯作者：
Jorgensen, Arne Lund
中科院分区：
文献类型：
--
作者：
Jakobsen, Jannik Ejnar;Johansen, Marianne G.;Schmidt, Mette;Dagnaes-Hansen, Frederik;Dam, Karen;Gunnarsson, Anders;Liu, Ying;Kragh, Peter M.;Li, Rong;Holm, Ida E.;Callesen, Henrik;Mikkelsen, Jacob Giehm;Nielsen, Anders Lade;Jorgensen, Arne Lund
使用位点特异性重组酶的靶向转基因是一种有吸引力的方法来创建转基因动物，因为它允许将转基因整合到预先选择的转录活性基因组位点。在这里，我们描述了重组酶介导的盒式交换（RMCE）在Göttingen迷你猪细胞中的应用，该细胞具有四个RMCE受体位点，每个位点包含一个由人类UbiC启动子驱动的绿色荧光蛋白（GFP）标记基因。4个RMCE受体位点相互独立分离，可在不同组织中确定表达谱。我们在具有所有四个受体位点的成纤维细胞中使用RMCE的微环，然后使用SCNT，生产了含有一个转录活性受体位点的转基因单拷贝的仔猪。该转基因由一个增强的人类UbiC启动子驱动的阿尔茨海默病致病基因PSEN1M146I的cDNA组成，其在各种组织中的表达谱与GFP标记基因相似。结果表明，RMCE可以在预先选择的转录活性受体位点上进行靶向转基因。本文的在线版本（doi:10.1007/s11248-012-9671-6）包含补充材料，仅供授权用户使用。
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. 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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影响因子：
4.8
作者：
Brevini, Tiziana A. L.;Pennarossa, Georgia;Gandolfi, Fulvio
通讯作者：
Gandolfi, Fulvio
影响因子：
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作者：
Carlson DF;Geurts AM;Garbe JR;Park CW;Rangel-Filho A;O'Grady SM;Jacob HJ;Steer CJ;Largaespada DA;Fahrenkrug SC
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作者：
Jakobsen, Jannik E.;Li, Juan;Nielsen, Anders Lade
通讯作者：
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