Transcriptional regulation of crb and mpp5 genes in nuclear space: Coupling nuclear polarization with cell polarization at the level of transcription

核空间中crb和mpp5基因的转录调控：在转录水平上将核极化与细胞极化耦合

• 批准号: 2221885
• 负责人: Xiangyun Wei
• 金额: $ 65万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2221885&HistoricalAwards=false
• 关键词: Transcriptional; regulation; crb; mpp5; genes

Differential gene expression is essential for normal tissue development and functions. Gene expression is regulated not only by cis- and trans-regulatory elements but also by the organization of the chromatin in the three-dimensional (3D) nuclear space. However, how the transcription of functionally-related genes is coordinated in 3D nuclear space in vivo is not clearly understood. This project will study how the differential transcription of the genes related to cell polarization and patterning are spatially coordinated in the nuclei of zebrafish retinal cells. The resultant polarization and patterning are important in light sensing in retinal cells. The significance of this study is also applicable to other tissues because polarity genes are required to be expressed in a coordinated fashion for proper tissue polarization, integrity, and function. The research will also contribute to the training of early career scientists and STEM workers through a structured mentoring program. The Crumbs and MPP5 genes and zebrafish retina form a unique system to study the roles of 3D nuclear organization in gene expression regulation. The Crumbs gene family has five members, and the MPP5 gene family has two members. Within the gene families, genes are differentially expressed in retinal cells; however, between the two families, some members are coexpressed in the same set of cell types for coordinated functions. The nuclei of zebrafish red, green, and blue cone photoreceptors (RGB cones) are almost twice as large as the nuclei of other retinal cells, offering the possibility of high-resolution 3D gene mapping. In addition, these nuclei are aligned in a mirror-symmetric pentameric pattern to allow defined spatial registration of gene positioning. This research will evaluate how in vivo nuclear positioning of these genes is correlated with their differential expression patterns by in situ hybridization and chromatin tagging. In addition, the roles of the enhancers of crb2b and mpp5b genes in gene positioning will be evaluated by genetic and transgenic approaches.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.

差异基因表达对于正常组织发育和功能是必不可少的。基因的表达不仅受顺式和反式调控元件的调控，而且还受染色质在三维（3D）核空间中的组织的调控。然而，功能相关基因的转录如何在体内3D核空间中协调尚不清楚。本计画将研究斑马鱼视网膜细胞核中与细胞极化及图案化相关基因的差异转录如何在空间上协调。由此产生的偏振和图案化在视网膜细胞中的光感测中是重要的。这项研究的意义也适用于其他组织，因为极性基因需要以协调的方式表达，以实现适当的组织极化、完整性和功能。该研究还将通过结构化的指导计划，为早期职业科学家和STEM工作者的培训做出贡献。 Crumbs和MPP 5基因和斑马鱼视网膜形成了一个独特的系统，以研究三维核组织在基因表达调控中的作用。Crumbs基因家族有五个成员，MPP 5基因家族有两个成员。在基因家族内，基因在视网膜细胞中差异表达;然而，在两个家族之间，一些成员在同一组细胞类型中共表达以协调功能。斑马鱼红、绿色和蓝色视锥光感受器（RGB视锥）的细胞核几乎是其他视网膜细胞细胞核的两倍，这为高分辨率3D基因定位提供了可能性。此外，这些细胞核以镜像对称的五聚体模式排列，以允许基因定位的定义的空间配准。本研究将通过原位杂交和染色质标记来评估这些基因在体内的核定位与其差异表达模式之间的关系。此外，crb 2b和mpp 5 b基因的增强子在基因定位中的作用将通过遗传和转基因方法进行评估。该奖项反映了NSF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。