CAREER: Priming CUREs to dissect how the DREAM complex protects cell identity

职业生涯：启动 CURE 来剖析 DREAM 复合物如何保护细胞身份

• 批准号: 2238540
• 负责人: Paul Goetsch
• 金额: $ 102.08万
• 依托单位: Michigan Technological University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2238540&HistoricalAwards=false
• 关键词: CAREER; Priming; CUREs; dissect; how

Cellular identity is controlled by protein networks that establish which genes are expressed in a cell. These protein networks react to internal and external signals and define how the cell functions within the context of a multicellular organism. A major obstacle in the field is that mechanisms that govern cell identity are remarkably complex, such that identifying the contributing factors remains a challenge. Identifying the missing components is critical for generating an integrated view of how specialized cell types develop. This project will fully integrate bioinformatic and experimental life science focused Course-based Undergraduate Research Experiences (CUREs) within the research activities to identity novel factors involved in establishing cell identity. Overall, the CURE efforts will offer undergraduate students equitable access to research and will help them develop the skills required to become competitive in their future life science careers.In Caenorhabditis elegans, the DREAM (for Dp, Retinoblastoma(Rb)-like, E2F, and MuvB) complex suppresses the differentiation programs in somatic (non-reproductive) cells, preventing them from activating the germ (reproductive) cell program, and also preserves the normal specification of vulval cell fate. DREAM represses genes through direct binding to their promoter regions, but no evidence indicates that the complex targets genes known to be associated with cell identity. These observations lead to the hypothesis that loss of the DREAM complex likely activates transcriptional components that drive breakdown of somatic cell identity. By creating improved reporter strains using CRISPR/Cas9-mediated genome editing in the laboratory with reverse genetic screens performed in the classroom, this proposal aims to reveal key components that contribute to normal germline and vulval development. In addition, the central role the DREAM complex plays in the regulation of cell identity in C. elegans offers an opportunity to explore how such developmental programs are conserved across multicellular organisms. Using bioinformatic conservation analyses and molecular genomics, this proposal will explore DREAM co-conservation with the intent of building a manual gene curation pipeline for identifying and annotating genes-of-interest in published but under-annotated genomes. Overall, this research will contribute to understanding the mechanisms by which transcriptional regulatory networks determine cellular and organismal diversity.This proposal was co-reviewed and will be co-funded by the Genetic Mechanisms Cluster in the Division of Molecular and Cellular Biosciences and by the Developmental Systems Program in the Division of Integrative Organismal Systems.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细胞身份是由蛋白质网络控制的，蛋白质网络决定了哪些基因在细胞中表达。这些蛋白质网络对内部和外部信号作出反应，并决定细胞如何在多细胞生物的背景下发挥作用。该领域的一个主要障碍是控制细胞身份的机制非常复杂，因此确定起作用的因素仍然是一个挑战。识别缺失的组件对于生成特化细胞类型如何发展的综合视图至关重要。该项目将在研究活动中充分整合生物信息学和以实验生命科学为重点的基于课程的本科生研究经验（CUREs），以确定与建立细胞身份相关的新因素。总体而言，CURE项目将为本科生提供公平的研究机会，并帮助他们发展在未来生命科学职业中具有竞争力所需的技能。在秀丽隐杆线虫中，DREAM （Dp，视网膜母细胞瘤样，E2F和MuvB）复合体抑制体细胞（非生殖）细胞的分化程序，阻止它们激活生殖（生殖）细胞程序，同时也保持了外阴细胞命运的正常规范。DREAM通过直接结合启动子区域抑制基因，但没有证据表明该复合物靶向已知与细胞身份相关的基因。这些观察结果导致了一个假设，即DREAM复合物的缺失可能激活了导致体细胞身份破坏的转录成分。通过在实验室中使用CRISPR/ cas9介导的基因组编辑创建改进的报告菌株，并在课堂上进行反向遗传筛选，该提案旨在揭示有助于正常种系和外阴发育的关键成分。此外，DREAM复合体在秀丽隐杆线虫细胞身份调节中的核心作用为探索这种发育程序如何在多细胞生物中保守提供了机会。利用生物信息学保护分析和分子基因组学，本提案将探索DREAM共同保护，目的是建立一个人工基因管理管道，用于识别和注释已发表但未注释的基因组中的感兴趣基因。总的来说，这项研究将有助于理解转录调控网络决定细胞和有机体多样性的机制。该提案由分子和细胞生物科学部的遗传机制组和综合有机体系统部的发育系统项目共同审查并共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。