Genetic interaction screen to analyze Robo signaling

用于分析 Robo 信号传导的遗传相互作用屏幕

基本信息

  • 批准号:
    7409740
  • 负责人:
  • 金额:
    $ 15.05万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2005
  • 资助国家:
    美国
  • 起止时间:
    2005-07-15 至 2010-04-30
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Slit-Robo signaling is 1 of the main axon guidance signaling pathways that act to shape the circuitry of the nervous system. Recent work shows the critical role of Slit-Robo signaling in the development of the visual system and spinal cord in model organisms, and the corticospinal tract in humans. However, only a few effector genes have been implicated in this signaling pathway, and none of these effector genes have been studied genetically in vertebrates. In addition, it is poorly understood which structural domains of Robo receptors are critical for their in vivo function. This proposal will use a forward genetic screen searching for interactions (noncomplementation) with the zebrafish astray (robo2) mutant. By screening individual F1 larvae labeled with a GFP transgene, we will be able to screen 75,000 genomes in 3 years. A pilot screen of 5700 genomes has already yielded 12 putative mutants. This screen will (1) identify mutants in genes in the Slit-Robo pathway, then (2) genetically map these mutants and (3) conduct a preliminary phenotypic assessment analyzing multiple axon pathways. We expect to find 3 classes of mutants: (I) new mutant alleles of robo2, which we will identify and sequence; (II) temperature-sensitive robo2 alleles, which we will identify and characterize; and (III) mutations in genes that show strong genetic interactions with robo2, such as slit ligands or downstream effectors, which we will genetically map and analyze phenotypically. The zebrafish system offers unique advantages for studying vertebrate axon guidance, including the ability to trace the trajectories of single axons both in fixed tissue and using timelapse movies; the ability to generate temperature-sensitive alleles in a vertebrate; and the ability to carry out forward-genetic screens. The robo2 alleles isolated in this screen will be valuable for understanding the timing of Robo2 function (temperature-sensitive alleles) and the protein domains necessary for Robo2 signaling (missense or nonsense alleles). The interacting genes isolated will define a set of genes in the Robo2 signaling pathway, and allow us to study their function in vivo.
描述(由申请人提供):Slit-Robo 信号传导是主要的轴突引导信号传导途径之一,其作用是塑造神经系统的电路。最近的研究表明,Slit-Robo 信号在模型生物的视觉系统和脊髓以及人类皮质脊髓束的发育中发挥着关键作用。然而,只有少数效应基因与该信号通路有关,并且这些效应基因都没有在脊椎动物中进行过遗传学研究。此外,人们对 Robo 受体的哪些结构域对其体内功能至关重要尚知之甚少。该提案将使用正向遗传筛选来寻找与斑马鱼迷途 (robo2) 突变体的相互作用(非互补)。通过筛选带有 GFP 转基因标记的 F1 幼虫个体,我们将能够在 3 年内筛选 75,000 个基因组。对 5700 个基因组的初步筛选已经产生了 12 个假定的突变体。该筛选将 (1) 识别 Slit-Robo 通路中基因的突变体,然后 (2) 对这些突变体进行基因图谱分析,以及 (3) 对多个轴突通路进行初步表型评估。我们期望找到 3 类突变体:(I) robo2 的新突变等位基因,我们将对其进行识别和测序; (II) 对温度敏感的 robo2 等位基因,我们将对其进行识别和表征; (III) 与 robo2 表现出强烈遗传相互作用的基因突变,例如狭缝配体或下游效应子,我们将对它们进行基因图谱和表型分析。斑马鱼系统为研究脊椎动物轴突引导提供了独特的优势,包括能够在固定组织中和使用延时电影追踪单个轴突的轨迹;在脊椎动物中产生温度敏感等位基因的能力;以及进行正向遗传筛选的能力。此筛选中分离的 robo2 等位基因对于了解 Robo2 功能的时间(温度敏感等位基因)和 Robo2 信号传导所需的蛋白质结构域(错义或无义等位基因)非常有价值。 The interacting genes isolated will define a set of genes in the Robo2 signaling pathway, and allow us to study their function in vivo.

项目成果

期刊论文数量(0)
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Chi-Bin Chien其他文献

Chi-Bin Chien的其他文献

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{{ truncateString('Chi-Bin Chien', 18)}}的其他基金

Genetic and imaging tools to visualize neuronal subsets in developing zebrafish
遗传和成像工具可可视化发育​​中的斑马鱼的神经元亚群
  • 批准号:
    7937648
  • 财政年份:
    2010
  • 资助金额:
    $ 15.05万
  • 项目类别:
PATTERNING OF DORSAL RETINA
背侧视网膜的图案化
  • 批准号:
    7193072
  • 财政年份:
    2006
  • 资助金额:
    $ 15.05万
  • 项目类别:
CONFOCAL MICROSCOPE FOR CORE IMAGING FACILITY: MUSCLE, LIMB DVMT, & CELL BIOL
用于核心成像设备的共焦显微镜:肌肉、肢体 DVMT、
  • 批准号:
    7166339
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
Genetic interaction screen to analyze Robo signaling
用于分析 Robo 信号传导的遗传相互作用屏幕
  • 批准号:
    7217454
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
Genetic interaction screen to analyze Robo signaling
用于分析 Robo 信号传导的遗传相互作用屏幕
  • 批准号:
    7093146
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
Genetic interaction screen to analyze Robo signaling
用于分析 Robo 信号传导的遗传相互作用屏幕
  • 批准号:
    6987375
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
CONFOCAL MICROSCOPE FOR CORE IMAGING FACILITY: ZEBRAFISH & C ELEGANS RESEARCH
用于核心成像设施的共焦显微镜:斑马鱼
  • 批准号:
    7166338
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
CONFOCAL MICROSCOPE FOR CORE IMAGING FACILITY: AIDS
用于核心成像设施的共焦显微镜:艾滋病
  • 批准号:
    7166337
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
Confocal Microscope for Core Imaging Facility
用于岩心成像设施的共焦显微镜
  • 批准号:
    6877361
  • 财政年份:
    2005
  • 资助金额:
    $ 15.05万
  • 项目类别:
RETINAL AXON PATHFINDING MUTANTS IN ZEBRAFISH
斑马鱼视网膜轴突寻路突变体
  • 批准号:
    6350905
  • 财政年份:
    2000
  • 资助金额:
    $ 15.05万
  • 项目类别:

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