Biochemistry in situ to determine inheritance of RNA-protein complexes

项目摘要 这项提议的总体目标是阐明RNA-蛋白质复合体是如何组装成 在动物世代之间传递信息。哪些RNA结合蛋白是遗传的？哪一个 细胞会遗传这些蛋白质吗？遗传需要哪些蛋白质属性？回答这些问题 这些问题需要开发新的原位生化方法来标记和跟踪特定的 RNA结合蛋白。该实验室使用线虫模型生物体，现代基因编辑 用于标记和跟踪生殖细胞RNA-蛋白质凝聚物的方法和较新的成像标签 动物体内的成分。这一策略需要大量的基因编辑和分子 进行基因筛查，以识别具有适当基因修改的动物。本副刊 请求部分支持以购买共享使用、凝胶和免疫印迹成像系统。这个 设备将被用来帮助识别具有正确基因类型、基因修改、 和蛋白质的表达。将提供机构支持，使更多人能够利用 科学界。成像器将能够有效地产生研究所需的动物 RNA结合蛋白在细胞和后代中的遗传。

Project Summary The overall objective of this proposal is to elucidate how RNA-protein complexes assemble to transmit information across animal generations. Which RNA binding proteins are inherited? Which cells inherit these proteins? What protein attributes are required for inheritance? Answering these questions require the development of new in situ biochemical methods to label and track specific RNA binding proteins. The lab uses the C. elegans model organism, modern gene editing methods, and newer imaging tags to label and track germ cell RNA-protein condensate components in animals. The strategy requires a significant amount of gene editing and molecular genetic screening to identify animals with the proper genetic modifications. This supplement requests partial support to purchase a shared-use, gel and immunoblot imaging system. The equipment will be used to help identify animals with the correct genotypes, gene modifications, and protein expression. Institutional support will be provided to enable additional use by the science community. The imager will enable efficient generation of animals necessary to study RNA binding protein inheritance in cells and progeny.