A novel gene trapping for identifying genes expressed under the control of specific transcription factors
A novel gene trapping for identifying genes expressed under the control of specific transcription factors
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DOI：
10.1016/j.bbrc.2007.06.161
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发表时间：
2007-09-14
影响因子：
3.1
通讯作者：
Asano, Masahide
中科院分区：
文献类型：
--
作者：
Naruse, Chie;Fukusumi, Yoshiyasu;Asano, Masahide
基因捕获法是一种鉴定新基因和分析其功能的有效方法。然而，很难根据它们的功能选择被捕获的基因。为了鉴定在中胚层形成中重要的转录因子调控的基因，我们通过感染表达Pax1、Brachyury和Foxa2的腺病毒载体选择了被困的ES克隆。在366个被捕获的基因中，在第一次筛选中有7个似乎是由这些转录因子控制的。用5' RACE技术对捕获的基因进行鉴定，北方印迹分析表明，3个捕获基因的表达受这些转录因子的调控。Cx43和HPI γ的表达模式暗示了它们在脊索和神经管形成中与Foxa2的功能关系。此外，Wtap突变小鼠来自被困的克隆表现出缺陷的中内胚层形成。我们的研究结果表明，被困ES克隆可以有效地选择使用转录因子。(c)2007爱思唯尔公司All rights reserved.
Gene trapping is a powerful method for identifying novel genes and for analyzing their functions. It is, however, difficult to select trapped genes on the basis of their function. To identify genes regulated by transcription factors that are important in the mesodermal formation, we selected trapped ES clones by infection of adenoviral vectors expressing Pax1, Brachyury, and Foxa2. Among 366 trapped genes, seven seemed to be controlled by these transcription factors in the first screening. The trapped genes were identified by 5' RACE, and a Northern blotting revealed that expressions of three trapped genes were regulated by these transcription factors. Expression patterns of Cx43 and HPI gamma implicated their functional relationships to Foxa2 in the formation of the notochord and the neural tube. Furthermore, Wtap mutant mice derived from the trapped clone showed defects in the mesendoderm formation. Our results indicate that trapped ES clones could be selected effectively using transcription factors. (c) 2007 Elsevier Inc. All rights reserved.