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

细胞特性由蛋白质网络控制，蛋白质网络确定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做出反应，并定义细胞如何在多细胞有机体中发挥作用。该领域的一个主要障碍是，管理细胞身份的机制非常复杂，因此识别促成因素仍然是一个挑战。识别缺失的成分对于生成专业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生命科学实验为重点的以课程为基础的本科生研究经验(CURE)，以确定建立细胞身份所涉及的新因素。总体而言，治疗努力将为本科生提供平等的研究机会，并将帮助他们发展在未来生命科学职业生涯中变得有竞争力所需的技能。在秀丽线虫中，梦想(对于DP、视网膜母细胞瘤(RB)样、E2F和MuvB)复合体抑制体细胞(非生殖)细胞的分化程序，阻止他们激活生殖细胞(生殖)计划，并保持外阴细胞命运的正常规格。Dream通过直接与基因的启动子区域结合来抑制基因，但没有证据表明这种复合体针对已知的与细胞身份相关的基因。这些观察结果导致了一种假设，即梦复合体的丧失可能激活了导致体细胞身份崩溃的转录成分。通过在实验室使用CRISPR/Cas9介导的基因组编辑创建改进的报告菌株，并在教室中进行反向遗传筛选，该提案旨在揭示有助于正常生殖系和外阴发育的关键成分。此外，梦复合体在线虫细胞身份调控中发挥的核心作用为探索这种发育程序如何在多细胞生物体中保守提供了机会。利用生物信息学保守分析和分子基因组学，该提案将探索DREAM共同保守，目的是建立一个手动基因管理管道，用于识别和注释已发表但注释不足的基因组中感兴趣的基因。总体而言，这项研究将有助于理解转录调控网络决定细胞和生物多样性的机制。这项提议得到了共同审查，并将由分子和细胞生物科学部门的遗传机制集群和综合组织系统部门的发展系统计划共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.