Analysis of mouse Cdh6 gene regulation by transgenesis of modified bacterial artificial chromosomes
Analysis of mouse Cdh6 gene regulation by transgenesis of modified bacterial artificial chromosomes
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DOI：
10.1016/j.ydbio.2007.12.011
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发表时间：
2008-03-15
影响因子：
2.7
通讯作者：
Krumlauf, Robb
中科院分区：
文献类型：
--
作者：
Inoue, Takayashi;Inoue, Yukiko U.;Krumlauf, Robb
经典的钙粘蛋白是细胞粘附分子，其表达模式与其在形态发生过程中的不同功能相关。钙粘蛋白基因座的大尺寸和复杂性使得研究控制其时空表达模式的顺式调节模块的组织成为一个挑战。为此，我们利用细菌人工染色体（BAC）含有Cdh 6基因，小鼠II型经典钙粘蛋白，系统地确定顺式调控模块，管理其表达。通过将lacZ报告基因插入Cdh 6 BAC中，并通过在转基因小鼠中检查的同源重组或转座子插入产生一系列修饰的变体，我们确定了一系列有助于基因特异性调控的基因组区域。这些区域的跨度类似于Cdh 6转录起始位点上游161-kb和下游186-kb之间的基因座的350 kb。在早期发育阶段，不同的模块独立地调节Cdh 6的隔室表达（即前脑、后脑菱形节和脊髓）和/或细胞系特异性表达模式（即神经嵴亚群，如施旺细胞）。关于神经嵴细胞中表达的调控，我们发现不同的区域对表达的不同方面有贡献，并且已经鉴定了一个短的79-bp区域，一旦细胞从神经管迁移，该区域就参与调控细胞中的表达。这些结果构建了Cdh 6顺式调节模块复杂组织的图片，并强调了有助于其在早期小鼠胚胎发育期间动态表达的多种输入。(C)2008年爱思唯尔公司All rights reserved.
Classic cadherins are cell adhesion molecules whose expression patterns are dynamically modulated in association with their diverse functions during morphogenesis. The large size and complexity of cadherin loci have made it a challenge to investigate the organization of cis-regulatory modules that control their spatiotemporal patterns of expression. Towards this end, we utilized bacterial artificial chromosomes (BACs) containing the Cdh6 gene, a mouse type II classic cadherin, to systematically identify cis-regulatory modules that govern its expression. By inserting a lacZ reporter gene into the Cdh6 BAC and generating a series of modified variants via homologous recombination or transposon insertions that have been examined in transgenic mice, we identified an array of genomic regions that contribute to specific regulation of the gene. These regions span similar to 350 kb of the locus between 161-kb upstream and 186-kb downstream of the Cdh6 transcription start site. Distinct modules independently regulate compartmental expression (i.e. forebrain, hindbrain rhombomeres, and spinal cord) and/or cell lineage-specific expression patterns (i.e. neural crest subpopulations such as Schwann cells) of Cdh6 at the early developmental stages. With respect to regulation of expression in neural crest cells, we have found that distinct regions contribute to different aspects of expression and have identified a short 79-bp region that is implicated in regulating expression in cells once they have emigrated from the neural tube. These results build a picture of the complex organization of Cdh6 cis-regulatory modules and highlight the diverse inputs that contribute to its dynamic expression during early mouse embryonic development. (C) 2008 Elsevier Inc. All rights reserved.