Regulatory sequences on the human villin gene trigger the expression of a reporter gene in a differentiating HT29 intestinal cell line.
Regulatory sequences on the human villin gene trigger the expression of a reporter gene in a differentiating HT29 intestinal cell line.
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人绒毛蛋白基因上的调控序列触发分化的 HT29 肠细胞系中报告基因的表达。
DOI：
10.1016/s0021-9258(18)82141-x
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
E. Pringault
中科院分区：
文献类型：
--
作者：
S. Robine;C. Sahuquillo;D. Louvard;E. Pringault
为了开发组织特异性靶向小肠和大肠粘膜未成熟和分化的上皮细胞中基因表达的分子工具，我们分离了人绒毛蛋白基因的2-kb 5 '侧翼区。该区域包含许多短序列，这些短序列在分化的肠上皮细胞中表达的基因的其他组织特异性启动子中是保守的。该DNA片段促进绒毛膜阳性肠、肾和肝癌细胞系中荧光素酶报告基因的转录和表达，但不促进绒毛膜阴性角质形成细胞系中荧光素酶报告基因的转录和表达。表达模式对应于内源基因的表达模式，表明该序列可以指导甜菜碱特异性转录。在分化的HT 29肠细胞系中，报告基因的表达在未分化的细胞中已经是可检测的，并且当诱导终末分化时显著增加。因此，如先前对于内源基因所报道的，绒毛蛋白基因的分离的5 '侧翼区对已知刺激这些培养的上皮肠细胞的终末分化的条件有积极响应。报道的结果表明，该基因组片段包含足够的调控元件，以概括绒毛启动子在肠分化过程中的表达模式。
To develop a molecular tool for tissue-specific targeting of gene expression in immature and differentiated epithelial cells of the small and large intestinal mucosa, we have isolated the 2-kb 5‘-flanking region of the human villin gene. This region contains numerous short sequences that are conserved among other tissue-specific promoters of genes expressed in differentiated enterocytes. This DNA fragment promotes the transcription and expression of the luciferase reporter gene in villin-positive intestinal, renal, and hepatoma cell lines but not in a villin-negative keratinocyte cell line. The pattern of expression corresponds that of the endogenous gene, indicating that this sequence can direct intestine-specific transcription. In the differentiating HT29 intestinal cell line, expression of the reporter gene is already detectable in undifferentiated cells, and dramatically increases when terminal differentiation is induced. Thus, as previously reported for the endogenous gene the isolated 5‘-flanking region of the villin gene responds positively to conditions known to stimulate terminal differentiation of these cultured epithelial intestinal cells. The reported results indicate that this genomic fragment contains sufficient regulatory elements to recapitulate the expression pattern of the villin promoter during intestinal differentiation.