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Dre 重组酶与 Cre 一样,是大肠杆菌、哺乳动物细胞和小鼠中一种高效的位点特异性重组酶。

Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice.

机构信息

Center for Regenerative Therapies Dresden, BioInnovationsZentrum Technische Universitaet Dresden, Am Tatzberg 47, 01307 Dresden, Germany.

出版信息

Dis Model Mech. 2009 Sep-Oct;2(9-10):508-15. doi: 10.1242/dmm.003087. Epub 2009 Aug 19.

Abstract

Tyrosine site-specific recombinases (SSRs) including Cre and FLP are essential tools for DNA and genome engineering. Cre has long been recognized as the best SSR for genome engineering, particularly in mice. Obtaining another SSR that is as good as Cre will be a valuable addition to the genomic toolbox. To this end, we have developed and validated reagents for the Dre-rox system. These include an Escherichia coli-inducible expression vector based on the temperature-sensitive pSC101 plasmid, a mammalian expression vector based on the CAGGs promoter, a rox-lacZ reporter embryonic stem (ES) cell line based on targeting at the Rosa26 locus, the accompanying Rosa26-rox reporter mouse line, and a CAGGs-Dre deleter mouse line. We also show that a Dre-progesterone receptor shows good ligand-responsive induction properties. Furthermore, we show that there is no crossover recombination between Cre-rox or Dre-loxP. Hence, we add another set of efficient tools to the genomic toolbox, which will enable the development of more sophisticated mouse models for the analysis of gene function and disease.

摘要

酪氨酸位点特异性重组酶(SSR)包括 Cre 和 FLP,是 DNA 和基因组工程的重要工具。Cre 长期以来一直被认为是基因组工程的最佳 SSR,特别是在小鼠中。获得另一种与 Cre 一样好的 SSR 将是基因组工具包的宝贵补充。为此,我们开发并验证了 Dre-rox 系统的试剂。这些试剂包括基于温度敏感型 pSC101 质粒的大肠杆菌诱导表达载体、基于 CAGGs 启动子的哺乳动物表达载体、基于靶向 Rosa26 基因座的 rox-lacZ 报告胚胎干细胞(ES)细胞系、伴随的 Rosa26-rox 报告小鼠系,以及 CAGGs-Dre 缺失小鼠系。我们还表明,Dre-孕激素受体表现出良好的配体反应诱导特性。此外,我们表明 Cre-rox 或 Dre-loxP 之间没有交叉重组。因此,我们为基因组工具包添加了另一组高效工具,这将使我们能够开发更复杂的小鼠模型,用于分析基因功能和疾病。

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