Alternative splicing and nonsense-mediated mRNA decay in neural development

神经发育中的选择性剪接和无义介导的 mRNA 衰减

• 批准号: 8878168
• 负责人: Sika Zheng
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-05 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8878168
• 关键词: Advisory Committees; Alternative Splicing; Area; Award; Binding; Binding Sites; Bioinformatics; Brain; Complement; Coupling; DNA Sequence Alteration; Data; Data Set; Defect; Development; Disease; Excitatory Synapse; Gene Expression; Genetic; Glutamate Receptor; Goals; High-Throughput Nucleotide Sequencing; Human; Immunoprecipitation; In Vitro; Knockout Mice; Knowledge; Lead; Light; Mammalian Cell; Mediating; Mental disorders; Mentors; Messenger RNA; MicroRNAs; Mission; Molecular Biology; Morphology; Nervous system structure; Neurons; Neurosciences; Outcome; Output; Pathogenesis; Pathway interactions; Phase; Physiological; Positioning Attribute; Proteins; Public Health; Quality Control; RNA; RNA Binding; RNA Processing; Regulation; Regulatory Pathway; Research; Research Project Grants; Rodent; Role; Scaffolding Protein; Specificity; Synapses; Techniques; Terminator Codon; Testing; Therapeutic Intervention; Training; Transcript; Transcriptional Regulation; Translations; Vertebral column; Yang; base; career development; collaborative environment; crosslink; crosslinking and immunoprecipitation sequencing; density; disability; experience; genome-wide; genome-wide analysis; in vivo; mRNA Decay; mRNA Transcript Degradation; mouse model; neurodevelopment; neuron development; neuropsychiatry; novel; postsynaptic; postsynaptic density protein; premature; research study; skills; tool; transcriptome sequencing

PROJECT SUMMARY Defects in the nonsense-mediated mRNA decay (NMD) pathway have recently been implicated in multiple neuropsychiatric diseases. However, this pathway has not been well studied in the brain and disruption of NMD in neurons has not been characterized. We recently discovered a novel regulatory role of the NMD pathway in the nervous system. Through coupling to alternative splicing regulation, NMD controls the expression level of postsynaptic density protein 95 (PSD-95), an essential scaffold protein of the excitatory PSD. This mechanism of regulation is similar to the output of transcriptional control, but it is unclear what other natural neuronal transcripts are regulated by NMD. My long term goal is to understand the role of NMD- mediated regulation in the nervous system, and how errors in NMD can lead to neuropsychiatric disorders. The objective of this proposal is to characterize fundamental developmental defects resulting from deficiencies in the NMD pathway and systematically identify physiological NMD substrates in rodent neurons. I have already generated an NMD deficient mouse model and developed various techniques for this proposal. Guided by strong preliminary data, I will pursue two specific aims: 1) Genome-wide identification of NMD targets important for neuronal development; 2) Characterize the physiological requirement for the NMD pathway during development. Based on preliminary data, two important aspects of synapse development, spine morphology and synaptic expression of glutamate receptors, will be examined thoroughly in NMD deficient neurons both in vivo and in vitro. Identification of natural NMD substrates in neurons will shed light on how frequently NMD is used as an active regulatory pathway of gene expression rather than a passive quality-control mechanism. Knowledge about the genetic roles and specific targets of NMD in brain provides a novel perspective for understanding the pathogenesis of various neuropsychiatric diseases and may help point to the common targets/pathways for therapeutic intervention. I have a background in both cellular neuroscience and RNA molecular biology. I am in a unique position to contribute to the proposed research area. To further prepare myself for my long-term research goal, I plan to seek training that will complement my existing technical skills and further develop my professional skills. UCLA has a highly collaborative environment ideal to this project and for me to achieve these goals. My mentor, Dr. Douglas Black, is one of the world-class experts in alternative splicing regulation. I also have an advisory committee that consists of Dr. Kelsey Martin, Dr. Tom O'Dell, and Dr. William Yang and provide complementary inputs on neuroscience. My mentor team has a detailed plan to facilitate my research progress and scientific career development. In summary, my educational and research experience together with a strong and supportive mentoring team make me an ideal candidate for this research project and the K99/R00 award.

项目摘要 无义介导的mRNA衰变（NMD）途径中的缺陷最近被牵连到多个 神经精神疾病然而，这种途径在大脑中还没有得到很好的研究， 神经元中的NMD尚未被表征。我们最近发现了NMD的一个新的调节作用 神经系统中的通路。通过与可变剪接调控偶联，NMD控制了 突触后密度蛋白95（PSD-95）的表达水平，PSD-95是兴奋性突触后膜的一种重要支架蛋白。 私营部门司这种调控机制类似于转录控制的输出，但还不清楚其他什么机制。 天然神经元转录物受NMD调节。我的长期目标是了解NMD的作用， 介导的调节在神经系统中，以及如何错误的NMD可以导致神经精神疾病。的 该提案的目的是描述由于以下方面的缺陷而导致的基本发育缺陷： NMD途径，并系统地鉴定啮齿动物神经元中生理NMD底物。我已经 产生了NMD缺陷小鼠模型，并为该提议开发了各种技术。指导 在获得大量初步数据后，我将追求两个具体目标：1）全基因组识别NMD目标的重要性 2）表征NMD通路在神经元发育期间的生理需求; 发展根据初步数据，突触发育的两个重要方面， 和谷氨酸受体的突触表达，将在NMD缺陷神经元中进行彻底检查， 体内和体外。神经元中天然NMD底物的鉴定将揭示NMD在神经元中的发生频率。 用作基因表达的主动调节途径，而不是被动的质量控制机制。 对NMD在脑中的遗传作用和特异性靶点的了解为NMD的研究提供了新的视角。 了解各种神经精神疾病的发病机制，并可能有助于指出共同的 治疗干预的目标/途径。 我有细胞神经科学和RNA分子生物学的背景。我处于一个独特的位置， 为拟议的研究领域做出贡献。为了进一步为我的长期研究目标做好准备，我计划 寻求能够补充我现有技术技能并进一步发展我的专业技能的培训。UCLA 有一个高度协作的环境，对这个项目和我实现这些目标非常理想。我的导师，博士。 道格拉斯布莱克是世界级的可变剪接调控专家之一。我还有一个建议 委员会由凯尔西马丁博士、汤姆奥戴尔博士和威廉杨博士组成， 对神经科学的补充投入。我的导师团队有一个详细的计划，以促进我的研究进展 科学的职业发展。总之，我的教育和研究经历以及 强大而支持的指导团队使我成为这个研究项目和K99/R 00的理想候选人 奖