RUI: Functional analysis of the exon junction complex in planarian stem cells and regeneration

RUI：涡虫干细胞外显子连接复合物的功能分析和再生

• 批准号: 1656793
• 负责人: Jason Pellettieri
• 金额: $ 47.44万
• 依托单位: Keene State College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1656793&HistoricalAwards=false
• 关键词: RUI; Functional; analysis; exon; junction

Non-Technical Paragraph: Many organisms, including humans, have some capacity to regenerate lost or damaged body parts following physical injury or disease. However, the extent of this ability varies considerably from one species to the next, and the biological mechanisms accounting for this variability are not well understood. This project will study genes that play a key role in the remarkable regenerative abilities of aquatic flatworms called planarians, animals that can re-form entire individuals from tiny body fragments in just over a week. Completion of the proposed objectives will advance our knowledge of how the large stem cell population that drives this process is regulated. Most research activities will be carried out by undergraduate students at Keene State College, providing valuable preparation for post-collegiate professional training and careers in the life sciences. Together with science outreach activities involving high school students, this will enhance New Hampshire's efforts to develop a technically proficient workforce crucial to further growth of the state's biotechnology and high-tech manufacturing industries.Technical Paragraph: This project will characterize post-transcriptional regulatory mechanisms governing gene expression in adult stem cells, using the planarian flatworm Schmidtea mediterranea as a model organism. Specifically, students will complete a functional analysis of the exon junction complex (EJC), a key regulator of RNA biochemistry with potentially conserved, yet poorly understood roles in stem cell biology. Prior work has established functions for the EJC in splicing, export of spliced mRNAs from the nucleus, translational control, and nonsense-mediated mRNA decay. The EJC has been linked to some gene-specific regulatory mechanisms during development; however, its biochemical function(s) and presumed RNA target(s) in self-renewing cell types in adult organisms are largely unknown. The research objectives of the current study will build upon preliminary data showing that the EJC is required for stem cell maintenance in planarians by: 1) Using RNAi and in situ hybridization to characterize the function and expression of EJC subunits during tissue homeostasis and regeneration; and 2) Combining candidate-gene and unbiased RNA-Seq approaches to identify EJC target RNAs. Completion of this research will enhance our understanding of molecular mechanisms governing gene expression in adult stem cells during tissue renewal and repair. The PI will undertake this work entirely with undergraduate and high school students to enhance STEM educational efforts in New Hampshire.

非技术性段落：包括人类在内的许多生物体都有一定的能力在身体受伤或疾病后再生丢失或受损的身体部位。 然而，这种能力的程度从一个物种到下一个物种变化很大，并且解释这种变化的生物学机制还没有很好地理解。 该项目将研究在称为Planarians的水生扁虫的显着再生能力中发挥关键作用的基因，这种动物可以在一周多的时间内从微小的身体碎片重新形成整个个体。 完成拟议的目标将推进我们的知识，如何驱动这一进程的大型干细胞群体的调节。 大多数研究活动将由基恩州立学院的本科生进行，为大学后的专业培训和生命科学职业生涯提供宝贵的准备。 再加上涉及高中生的科学推广活动，这将加强新罕布什尔州的努力，以发展一个技术熟练的劳动力至关重要的国家的生物技术和高科技制造业的进一步增长。Technical Paragraph：本项目将特点转录后调节机制，在成人干细胞基因表达，使用作为模式生物的扁形虫Schmidtea mediterranea。 具体来说，学生将完成外显子连接复合物（EJC）的功能分析，这是RNA生物化学的关键调节因子，具有潜在的保守性，但在干细胞生物学中的作用知之甚少。 先前的工作已经确定了EJC在剪接、从细胞核输出剪接的mRNA、翻译控制和无义介导的mRNA衰变中的功能。 EJC在发育过程中与一些基因特异性调控机制有关;然而，其生化功能和成年生物体自我更新细胞类型中的假定RNA靶标在很大程度上是未知的。 本研究的研究目标将建立在初步数据的基础上，这些数据表明，EJC是涡虫干细胞维持所必需的：1）使用RNAi和原位杂交来表征组织稳态和再生过程中EJC亚基的功能和表达; 2）结合候选基因和无偏见的RNA-Seq方法来鉴定EJC靶RNA。 这项研究的完成将提高我们对组织更新和修复过程中成体干细胞基因表达的分子机制的理解。 PI将完全与本科生和高中生一起开展这项工作，以加强新罕布什尔州的STEM教育工作。