WINGED HELIX TRANSCRIPTION FACTOR FOXD3 IN NEURAL CREST DEV AND EVOLUTION

神经嵴发育和进化中的翼状螺旋转录因子 FOXD3

• 批准号: 7381542
• 负责人: MARK V REEDY
• 金额: $ 1.33万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381542
• 关键词: WINGED; HELIX; TRANSCRIPTION; FACTOR; FOXD3

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. During the past year my lab has focused on specific aim #1 and specific aim #3, experiment 2 of my grant. Specific aim #1 is to identify the regulatory elements driving neural crest-specific expression of the chicken homology of FoxD3 (cFoxd). To this end we cloned an approximately 3kb genomic DNA fragement 5' to cFoxD3, inserted it into the pBGZA reporter vector, and used in ovo electroporation to transfect the construct into the dorsal neural tube of stage 11 chicken embryos. This construct did not drive reporter gene expression the neural tube or neural crest cells, so this summer we will begin cloning larger 5' fragments and some 3' fragments to test. Specific aim #3, experiment 2 involves determining whether the regulatory elements from AmphifoxD are capable of driving reporter gene expression in the vertebrate neural tube and/or neural crest cells. Our data thus far indicate that they are not, suggesting that that the difference in expression of foxD3 between amphioxus and vertebrate embryos is due primarily to cis-regulatory factors rather than trans-regulatory factors. In addition to continuing our work on specific aim #3, we will begin work on specific aim #2 (generating constitutive activator and repressor forms of cFoxD3) and specific aim #3, experiment 1 (testing the ability of AmphifoxD to induce ectopic neural crest cells when expressed in the vertebrate neural tube). I will be doing this work with Rachel Patterson, a sophomore Biology major and my first INBRE scholar.

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。在过去的一年里，我的实验室一直专注于特定目标#1和特定目标#3，即我的资助的实验2。具体目的#1是确定驱动FoxD3 （cFoxd）神经冠特异性表达的调控元件。为此，我们克隆了一个约3kb的基因组DNA片段5'到cFoxD3上，将其插入pBGZA报告载体中，并利用卵细胞电穿孔将其转染到11期鸡胚胎的背神经管中。这种结构不能驱动神经管或神经嵴细胞的报告基因表达，因此今年夏天我们将开始克隆较大的5‘片段和一些3’片段进行测试。具体目标#3，实验2涉及确定来自AmphifoxD的调节元件是否能够驱动脊椎动物神经管和/或神经嵴细胞中的报告基因表达。到目前为止，我们的数据表明它们不是，这表明文昌鱼和脊椎动物胚胎之间foxD3表达的差异主要是由于顺式调节因子而不是反式调节因子。除了继续我们在特定目标#3上的工作外，我们将开始研究特定目标#2（生成cFoxD3的组成激活剂和抑制剂形式）和特定目标#3，实验1（测试AmphifoxD在脊椎动物神经管中表达时诱导异位神经嵴细胞的能力）。我将和Rachel Patterson一起做这项工作，她是生物学专业的大二学生，也是我的第一位INBRE学者。