Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction.
Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction.
复制标题
通过联合使用CRE介导的重组和四环素诱导，在小鼠中有条件和可诱导的转基因表达。
DOI：
10.1093/nar/gni051
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发表时间：
2005-03-22
影响因子：
14.9
通讯作者：
Nagy, Andras
中科院分区：
文献类型：
--
作者：
Belteki, Gusztav;Haigh, Jody;Kabacs, Nikolett;Haigh, Katharina;Sison, Karen;Costantini, Frank;Whitsett, Jeff;Quaggin, Susan E;Nagy, Andras
在这里，我们描述了一个三重转基因小鼠系统，它结合了任何cre转基因系的组织特异性和逆转录四环素反应因子(rtTA)/四环素反应元件（tet-O）驱动转基因的诱导性。为了确保在广泛的细胞类型中可靠地表达rtTA，我们将rtTA转基因靶向到ROSA26位点。然而，rtTA表达的条件是Cre重组酶介导的ROSA26位点STOP区域的切除。我们通过在成年小鼠胚胎发育和出生后诱导表达血管内皮生长因子（VEGF-A）来证明该技术的实用性。我们的成人诱导结果概括了几种不同的肝脏和免疫细胞病理表型与增加的全身VEGF-A蛋白水平相关。该系统将有助于研究基因，其中表达的时间控制是发现全谱功能所必需的。所提出的方法消除了为已经存在良好表征的Cre系的组织产生组织特异性rtTA转基因的需要。
Here we describe a triple transgenic mouse system, which combines the tissue specificity of any Cre-transgenic line with the inducibility of the reverse tetracycline transactivator (rtTA)/tetracycline-responsive element (tet-O)-driven transgenes. To ensure reliable rtTA expression in a broad range of cell types, we have targeted the rtTA transgene into the ROSA26 locus. The rtTA expression, however, is conditional to a Cre recombinase-mediated excision of a STOP region from the ROSA26 locus. We demonstrate the utility of this technology through the inducible expression of the vascular endothelial growth factor (VEGF-A) during embryonic development and postnatally in adult mice. Our results of adult induction recapitulate several different hepatic and immune cell pathological phenotypes associated with increased systemic VEGF-A protein levels. This system will be useful for studying genes in which temporal control of expression is necessary for the discovery of the full spectrum of functions. The presented approach abrogates the need to generate tissue-specific rtTA transgenes for tissues where well-characterized Cre lines already exist.