General and Conditional Replacement of Connexin43-Coding DNA by a lacZ Reporter Gene for Cell-Autonomous Analysis of Expression
General and Conditional Replacement of Connexin43-Coding DNA by a lacZ Reporter Gene for Cell-Autonomous Analysis of Expression
复制标题
用 lacZ 报告基因一般和有条件地替换 Connexin43 编码 DNA，用于细胞自主表达分析
DOI：
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发表时间：
2001
影响因子：
--
通讯作者：
K. Willecke
中科院分区：
文献类型：
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作者：
M. Theis;C. Mas;Britta Döring;O. Krüger;P. Herrera;P. Meda;K. Willecke
利用Cre/loxP系统，我们规避了Cx43基因缺失的早期产后死亡和可能的多效性影响，以确定Cx43在特定细胞类型中的表达和功能。一般或细胞类型特异性，cre介导的Cx43编码区封闭（即两侧有loxP位点）的缺失导致插入的lacZ报告基因在具有Cx43基因转录活性的细胞中被激活。根据lacZ在缺失小鼠中的表达推断，Cx43基因的转录活性不仅存在于已知表达Cx43的多种细胞类型中，还存在于胰腺管细胞、肠道血管细胞和骨骼肌中。cre介导的缺失仅限于特定的细胞类型，导致lacZ激活突出相应的表达Cx43的细胞亚群，如血管内皮细胞、肝管细胞和假定的神经嵴细胞，否则这些细胞会被邻近细胞中强烈的Cx43表达所掩盖。在表达Cx43的细胞类型中，固定的Cx43等位基因可作为Cre切除报告基因，用于表征Cre转基因。
Using the Cre/loxP system, we have circumvented early postnatal lethality and possible pleiotropic effects of general Cx43 gene deletion, in order to determine the expression and function of connexin43 (Cx43) in defined cell types. General or cell type-specific, Cre-mediated deletion of the floxed (i.e. flanked by loxP sites) Cx43-coding region led to activation of the inserted lacZ reporter gene in cells with transcriptional activity of the Cx43 gene. As deduced from lacZ expression in mice with general deletion, transcriptional activity of the Cx43 gene was not only found in a broad range of cell types known to a express Cx43, but also in pancreatic duct cells and vascular cells of the gut and skeletal muscle. Cre-mediated deletion restricted to defined cell types led to lacZ activation highlighting corresponding subsets of cells expressing Cx43, such as vascular endothelial cells, hepatic duct cells and putative neural crest cells, which were otherwise masked by strong Cx43 expression in neighbouring cells. In Cx43 expressing cell types, the floxed Cx43 allele was useful as a Cre-excision reporter for the characterization of Cre transgenes.
影响因子：
6.6
作者：
DECARVALHO, ACC;ROY, C;SPRAY, DC
通讯作者：
SPRAY, DC