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首页> 外文期刊>Biochemistry >Functional Characterization of Transgene Integration Patterns by Halo Fluorescence in Situ Hybridization: Electroporation versus Retroviral Infection
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Functional Characterization of Transgene Integration Patterns by Halo Fluorescence in Situ Hybridization: Electroporation versus Retroviral Infection

机译：卤素荧光在原位杂交中的转基因整合模式的功能表征：电穿孔与逆转录病毒感染

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摘要

Gene transfer techniques are essential for bioengineering and gene therapy. Retroviral vectors constitute an important tool in this context as integrase-catalyzed genomic anchoring is one of the best defined results of nonhomologous recombination, occurring at loci with favorable expression properties. One the basis of a retroviral expression cassette, this study focuses on differences regarding the genomic targets after its transfer by either retroviral infection or electroporation. Fluorescence in situ hybridization (FISH) of clones generated by infection revealed for the majority an association of the transgene with the nuclear matrix while this was true only for a minority of single copy clones if the same construct was transferred by electroporation. Our results demonstrate that the process of integration is distinct for both gene transfer routes an confirm that retroviral survival strategies favor integration next to scaffold/matrix attachment regions. These results may be one key to explain recent consequences of gene therapy trials that have led to the deregulation of endogenous genes.
机译：基因转移技术对于生物工程和基因治疗是必不可少的。逆转录病毒载体在这种情况下构成一个重要的工具，因为整合酶催化的基因组锚定是在具有有利表达性质的基因座中发生的非致素重组的最佳定义结果之一。该研究归一下逆转录病毒表达盒的基础，侧重于通过逆转录病毒感染或电穿孔转移后的基因组靶标的差异。通过感染产生的克隆的荧光杂交（鱼类）揭示了大多数转基因与核基质的结合，而当通过电穿孔转移相同的构建体，这对于少数单拷贝克隆，这是真的。我们的结果表明，两种基因转移路线都有明显的，确认逆转录病毒生存策略有利于脚手架/矩阵附件区域旁边的积分。这些结果可能是解释基因治疗试验的最近后果导致对内源基因进行放松的后果的一个关键。

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  • 来源
    《Biochemistry》 |2003年第23期|共9页
  • 作者

    S. Goetze; Y. Huesemann; A. Baer; J. Bode;

  • 作者单位

    GBF-German Research Center for Biotechnology/Epigenetic Regulation Mascheroder Weg 1 D-38124 Braunschweig Germany;

    GBF-German Research Center for Biotechnology/Epigenetic Regulation Mascheroder Weg 1 D-38124 Braunschweig Germany;

    GBF-German Research Center for Biotechnology/Epigenetic Regulation Mascheroder Weg 1 D-38124 Braunschweig Germany;

    GBF-German Research Center for Biotechnology/Epigenetic Regulation Mascheroder Weg 1 D-38124 Braunschweig Germany;

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  • 正文语种 eng
  • 中图分类 生物化学;
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