EAGER: In Vivo Detection of Protein-Protein Interactions

智力优势：分子和遗传学研究表明，蛋白质在多聚体复合物中起作用，以指导细胞事件，特别是转录因子，可以使用选择性相互作用来差异调节基因表达。 用于检测这些相互作用的现有方法在它们提供的信息方面是有限的，通常是因为检测方法最适合于在细胞培养物或单细胞生物中进行的研究。 该项目涉及开发一种变革性的方法来检测活体动物细胞核内转录因子之间的瞬时相互作用。 该项目将使用模式生物秀丽隐杆线虫，这是一种通常用于发育和遗传研究的小型多细胞蠕虫。 荧光成像将用于检测与核膜相连的转录因子与荧光标记配偶体之间的相互作用。 概念验证实验将与已知的互动者进行，一旦建立，该方法将被应用于发现新的互动伙伴。这项研究是新颖的，潜在的高意义，因为它将允许前所未有的在体内检测转录因子之间的相互作用，在整个生命周期的organis.Broader影响：本科生，硕士，博士生从格鲁吉亚州立大学，一个民族和种族多元化的城市机构，将参与这个项目的各个方面。 除了提供研究培训外，该项目还将为两名大学生提供国际研究经验。 该项目还将通过利文斯顿学院教师和学生的夏季研究经验以及格鲁吉亚州立大学教师对利文斯顿的学年讲座/访问，加强利文斯顿学院的研究基础设施。 此外，博士后研究员将获得在本科和研究生课堂上的监督和指导教学经验。

Intellectual Merit: Molecular and genetic studies have shown that proteins work in multimeric complexes to direct cellular events, and that transcription factors, in particular, can use selective interactions to differentially regulate gene expression. Existing methods for detecting these interactions are limited in the information that they provide, usually because the method of detection is most appropriate for studies done in cell culture or in single-celled organisms. This project involves developing a transformative method to detect transient interactions among transcription factors within the nuclei of the cells of living animals. The project will use the model organism, Caenorhabditis elegans, a small multicellular worm commonly used for developmental and genetic studies. Fluorescence imaging will be used detect interaction between a transcription factor tethered to the nuclear membrane and a fluorescently-labeled partner. Proof-of-concept experiments will be carried out with known interactors and,once established, the method will be applied to discover novel interaction partners. The research is novel and potentially of high significance because it would allow unprecedented in vivo detection of interaction between transcription factors throughout the life cycle of an organism.Broader Impacts: Undergraduate, Masters, and PhD students from Georgia State University, an ethnically and racially diverse urban institution, will participate in all aspects of this project. In addition to providing research training, this project will provide an international research experience for two college students. The project will also enhance the research infrastructure at Livingstone College, a historically black college, through a combination of summer research experiences for Livingston College faculty and students and academic year lectures/visits to Livingstone by Georgia State University faculty. Furthermore, a postdoctoral fellow will gain supervised and guided teaching experience in both undergraduate and graduate classrooms.