Pou4 Function in an Invertebrate Chordate
无脊椎动物脊索动物中的 Pou4 功能
基本信息
- 批准号:1557448
- 负责人:
- 金额:$ 30万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:2016
- 资助国家:美国
- 起止时间:2016-08-01 至 2020-07-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Transcription factors (TF) are proteins that control the expression of genes. This project uses a simple animal that is closely related to vertebrates like ourselves, the ascidian, to study a TF called Pou4 that is important for the development of sensory neurons, which include the hair cells of the inner ear. It was previously found that expressing ascidian Pou4 in ascidian cell types where it is not usually found converts those cells into sensory neurons. Likewise, forced expression of any of the three mouse Pou4 genes in the wrong ascidian cell types also converts those cells into sensory neurons. However, in mice, expressing Pou4 in the wrong cell type does not convert those cells into sensory neurons. The present project examines the mechanistic basis for the differences between the ascidian and mouse Pou4 genes in order to better understand evolutionary changes in development that give rise to new cell types like vertebrate sensory neurons. To answer these questions, the researchers are adapting a new gene editing system (CRISPR/Cas9) to the ascidian to decipher the mechanisms behind Pou4 function. Broader Impacts: San Diego State University ranks among the top US universities for economic and campus ethnic diversity. The proposed study will provide laboratory research opportunities for undergraduate and graduate students. To broaden the overall understanding of the function and experimental use of transgenic organisms to SDSU's larger undergraduate population, the researchers are working with the coordinator of their introductory cell and molecular biology courses to develop a relatively simple, "hands on" exercise where students can generate and assay transgenic ascidian embryos.In ascidians, the single Pou4 (CiPou4) transcription factor (TF) is a key regular of the PNS, and ectopic expression of this gene produces ectopic sensory neurons. In mammals, there are three Pou4s; all predominately expressed in sensory neurons. The investigators used RNA-SEQ to identify CiPou4 transcriptional targets and many are known to be expressed in vertebrate hair cells, suggesting a deep evolutionary linkage between these cells types. In mammals, ectopic expression of the hair cell Pou4F3 gene does not produce new hair cells; but when expressed in ascidians ectopic sensory neurons are generated. This observation suggests that while protein function is conserved, Pou4 transcriptional targets have been lost and/or are differentially regulated in vertebrates. Here we will use RNA-SEQ to determine if the mouse Pou4 TFs activate similar transcriptional targets compared to CiPou4. We will test if a CRISPR/Cas9-mediated CiPou4 knock-out can be rescued by the mouse Pou4 genes. This project is transformative because the ascidian larva provides a unique system to examine the evolution of TF function and the role this TF has played during the evolution of a novel vertebrate cell type, the sensory hair cell.
转录因子(TF)是控制基因表达的蛋白质。该项目使用一种与我们这样的脊椎动物密切相关的简单动物——海鞘,来研究一种名为 Pou4 的转录因子,它对于感觉神经元(包括内耳毛细胞)的发育非常重要。此前发现,在通常不存在的海鞘细胞类型中表达海鞘 Pou4 可以将这些细胞转化为感觉神经元。同样,在错误的海鞘细胞类型中强制表达三种小鼠 Pou4 基因中的任何一种也会将这些细胞转化为感觉神经元。然而,在小鼠中,在错误的细胞类型中表达 Pou4 不会将这些细胞转化为感觉神经元。本项目研究了海鞘和小鼠 Pou4 基因之间差异的机制基础,以便更好地理解发育过程中产生新细胞类型(如脊椎动物感觉神经元)的进化变化。为了回答这些问题,研究人员正在将一种新的基因编辑系统 (CRISPR/Cas9) 应用于海鞘,以破译 Pou4 功能背后的机制。更广泛的影响:圣地亚哥州立大学在经济和校园种族多样性方面位居美国顶尖大学之列。拟议的研究将为本科生和研究生提供实验室研究机会。为了扩大 SDSU 更大的本科生群体对转基因生物功能和实验用途的整体了解,研究人员正在与细胞和分子生物学导论课程的协调员合作,开发一种相对简单的“动手”练习,学生可以在其中生成并分析转基因海鞘胚胎。在海鞘中,单个 Pou4 (CiPou4) 转录因子 (TF) 是 PNS,该基因的异位表达会产生异位感觉神经元。在哺乳动物中,有 3 个 Pou4;全部主要在感觉神经元中表达。研究人员使用 RNA-SEQ 来鉴定 CiPou4 转录靶标,已知许多转录靶标在脊椎动物毛细胞中表达,表明这些细胞类型之间存在深刻的进化联系。在哺乳动物中,毛细胞Pou4F3基因的异位表达不会产生新的毛细胞;但当在海鞘中表达时,会产生异位感觉神经元。这一观察结果表明,虽然蛋白质功能是保守的,但 Pou4 转录靶点在脊椎动物中已经丢失和/或受到差异性调节。在这里,我们将使用 RNA-SEQ 来确定小鼠 Pou4 TF 是否激活与 CiPou4 相似的转录目标。我们将测试小鼠 Pou4 基因是否可以挽救 CRISPR/Cas9 介导的 CiPou4 敲除。该项目具有变革性,因为海鞘幼虫提供了一个独特的系统来检查 TF 功能的进化以及该 TF 在新型脊椎动物细胞类型(感觉毛细胞)进化过程中所发挥的作用。
项目成果
期刊论文数量(0)
专著数量(0)
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Robert Zeller其他文献
Idiopathic muscular hypertrophy of the esophagus: A case report
- DOI:
10.1007/bf01887509 - 发表时间:
1979-12-01 - 期刊:
- 影响因子:2.200
- 作者:
Robert Zeller;Robert McLelland;Beverly Myers;William M. Thompson - 通讯作者:
William M. Thompson
Robert Zeller的其他文献
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{{ truncateString('Robert Zeller', 18)}}的其他基金
The Interplay Between Notch Ligands is Critical for Epidermal Sensory Neuronal Specification and Spatial Patterning in the Ascidian Embryo
缺口配体之间的相互作用对于海鞘胚胎的表皮感觉神经元规格和空间模式至关重要
- 批准号:
1938531 - 财政年份:2020
- 资助金额:
$ 30万 - 项目类别:
Standard Grant
The regulation and function of miR-124 in ascidian neural development
miR-124在海鞘神经发育中的调控及功能
- 批准号:
0951347 - 财政年份:2010
- 资助金额:
$ 30万 - 项目类别:
Continuing Grant
CAREER: Development of the Ascidian Peripheral Nervous System
职业:海鞘周围神经系统的发育
- 批准号:
0347937 - 财政年份:2004
- 资助金额:
$ 30万 - 项目类别:
Continuing Grant
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