A specific promoter of the sensory cells of the inner ear defined by transgenesis
A specific promoter of the sensory cells of the inner ear defined by transgenesis
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DOI：
10.1093/hmg/10.15.1581
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发表时间：
2001-07-15
影响因子：
3.5
通讯作者：
Petit, C
中科院分区：
文献类型：
--
作者：
Boëda, B;Weil, D;Petit, C
迄今为止，还没有报道针对内耳感觉细胞（毛细胞）的特异性启动子序列。为了了解决定内耳毛细胞命运的分子机制，并以开发用于基因治疗和产生条件性敲除的有价值的工具为目标，我们开始寻找调节鼠Myo 7a和人MYO 7A基因表达的顺式作用DNA序列。这些基因编码非常规的肌球蛋白VIIA，其在毛细胞和一些其他上皮细胞中表达。我们产生了转基因小鼠品系，这些转基因小鼠在Myo 7a/MYO 7A启动子区和内含子1的几个5 ' -截短形式的控制下表达绿色荧光蛋白（GFP）报告基因。我们获得了GFP表达仅限于内耳、耳蜗和前庭毛细胞的转基因小鼠。启动子的连续缺失使我们能够定义在内含子1存在下足以将转基因表达靶向毛细胞的118 bp的最小序列。此外，从转基因中删除内含子1消除了毛细胞表达，从而表明内含子中存在强增强子。这是第一次报告的调控序列足以靶向表达的基因只在感觉细胞的内耳。这也为毛细胞转录组的分析开辟了可能性。
To date, no promoter sequence specific to the inner ear sensory cells (hair cells) has been reported. In an effort to understand the molecular mechanisms that determine hair cell fate in the inner ear, and with the goal of developing a valuable tool for gene therapy and for the generation of conditional knockouts, we initiated a search for cis-acting DNA sequences that regulate the expression of the murine Myo7a and human MYO7A genes. These genes encode the unconventional myosin VIIA which is expressed in hair cells and in some other epithelial cells. We generated lines of transgenic mice expressing the green fluorescent protein (GFP) reporter gene under the control of several 5 ' -truncated versions of the Myo7a/MYO7A promoter region and intron 1. We obtained transgenic mice with a GFP expression restricted to the hair cells of the inner ear, cochlea and vestibule. Successive deletions of the promoter allowed us to define a minimal sequence of 118 bp that is sufficient, in the presence of intron 1, to target the transgene expression to hair cells. In addition, the deletion of intron 1 from the transgenes abolished hair cell expression, thus indicating the presence of a strong enhancer in the intron. This is the first report of regulatory sequences sufficient to target the expression of a gene exclusively in sensory cells of the inner ear. It also opens up the possibility for the analysis of the hair cell transcriptome.