Identification of downstream signals for Pitx2a

Pitx2a 下游信号的识别

• 批准号: 6556774
• 负责人: QIZE WEI
• 金额: $ 17.83万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6556774
• 关键词: DNA binding protein; HeLa cells; RNA; biological signal transduction; cell growth regulation; cell motility; cell proliferation; developmental genetics; embryogenesis; gene induction /repression; genetic regulatory element; genetically modified animals; histogenesis; laboratory mouse; nucleic acid sequence; protein isoforms; protein structure function; tissue /cell culture; transcription factor

DESCRIPTION (provided by applicant): Pitx2, a bicoid-type homeodomain transcription factor, has been implicated in Rieger's syndrome in humans and plays an important role in mediating left-right development in mice, chickens, frogs and zebrafish. It is believed that cell proliferation, cell motility as well as changes in instructive signals for cell fate in local areas are important for directing left-right asymmetry during organogenesis. As a transcription factor, Pitx2 should activate and/or repress the transcription of its target genes to execute specific cellular functions. However, little is known about the downstream targets and signaling pathways for Pitx2. I propose in this study to search for the target genes regulated by Pitx2a (one of the Pitx2 isoforms) and the relevant signaling pathways. I will use an inducible HeLa cell line ectopically expressing Pitx2a, which is already available in the laboratory. This inducible cell line will allow me to trace the downstream signaling pathways for Pitx2a based on cell phenotype and to identify the target genes for Pitx2a by comparing the RNA isolated from cells under the induced and uninduced conditions. The transgenic mouse model will also be used to assess the importance of the putative downstream target genes in mediating the function of Pitx2a in mice. Identification of these downstream target genes and signaling pathways using these cell culture and mouse model systems should allow me to make progress towards understanding of the mechanism underlying the functions of Pitx2a as well as the missing links between Pitx2a expression and its roles in morphogenesis during embryonic development.

描述（由申请人提供）： Pitx2 是一种 Bicoid 型同源域转录因子，与人类 Rieger 综合征有关，并在介导小鼠、鸡、青蛙和斑马鱼的左右发育中发挥重要作用。 据信，细胞增殖、细胞运动以及局部区域细胞命运指导信号的变化对于引导器官发生过程中的左右不对称非常重要。 作为转录因子，Pitx2 应激活和/或抑制其靶基因的转录以执行特定的细胞功能。 然而，人们对 Pitx2 的下游靶点和信号通路知之甚少。 我建议在这项研究中寻找Pitx2a（Pitx2亚型之一）调控的靶基因和相关信号通路。 我将使用异位表达 Pitx2a 的诱导型 HeLa 细胞系，该细胞系已在实验室中可用。 这种诱导细胞系将使我能够根据细胞表型追踪 Pitx2a 的下游信号通路，并通过比较在诱导和非诱导条件下从细胞中分离的 RNA 来识别 Pitx2a 的靶基因。 转基因小鼠模型还将用于评估假定的下游靶基因在介导小鼠 Pitx2a 功能中的重要性。 使用这些细胞培养物和小鼠模型系统鉴定这些下游靶基因和信号通路应该使我能够在理解 Pitx2a 功能背后的机制以及 Pitx2a 表达及其在胚胎发育过程中形态发生中的作用之间缺失的联系方面取得进展。