Mouse Genetics and Cell Line Core

项目总结（见说明）： 在过去十年中，生物医学科学技术应用取得了巨大进展。其中更显着的成就是全基因组测序的出现，允许完整的知识的基因组DNA结构的inunierable生物和相关的分子生物学的进步，使这些新发现的基因组序列的全面操纵。随之而来的是，有能力提升对肾脏疾病的科学研究，并改进哺乳动物肾脏的体内系统。肾脏疾病的基础科学发现的验证现在取决于对基因工程小鼠模型和自然发生的人类样本的研究。耶鲁大学乔治M.奥布莱恩肾脏中心旨在减少障碍，促进基于小鼠的体内技术在肾脏疾病研究中的应用，并促进研究扩展到源自工程突变小鼠的基于细胞的离体系统。核心的具体目标是对细菌人工染色体（BAC）中的目标基因进行修饰，以用于转基因小鼠;从突变小鼠品系的特定肾单位片段中分离初级小管ceNs和条件永生化细胞系;促进产生条件敲除和敲入基因靶向策略和构建体，并为小鼠遗传学应用研究人员提供一般资源。该核心将与肾脏生理学和表型核心协调，以协助研究者将小鼠遗传服务与生理学研究结合起来。它将与人类遗传学和临床研究协调，将新的人类疾病基因发现与动物和细胞系模型相结合。该核心的小鼠模型和细胞系组分具有固有的协同作用，因为它允许研究者访问从定义的体内模型到具有相同遗传组成的离体细胞系模型的范围。核心现在增加了一个新的机制，细胞系生成的基础上，核心教师开发的技术，一个改进的BAC重组方法和一个新的服务，在条件敲除或敲入等位基因生成的基础上BAC技术。

PROJECT SUMMARY (See instructions): The past decade has seen quantum advances in the applications of technology to biomedical sciences. Among the more remarkable achievements has been the advent of whole genome sequencing allowing complete knowledge of the genomic DNA structure of innunierable organisms and the associated molecular biologic advances enabling comprehensive manipulation of these newly discovered genomic sequences. With this has come the ability to elevate scientific investigation of kidney disease and function to refined in vivo systems in the mammalian kidney. The validation of basic scientific discoveries in kidney disease now rests with studies in genetically engineered mouse models and naturally occurring human samples. The overarching objective of the Mouse Genetics and Cell Line Core in the Yale George M. O'Brien Kidney Center is to reduce barriers and facilitate application of in vivo mouse-based technologies to the study of kidney disease and to facilitate the extension of the studies to ex vivo cell-based systems derived from engineered mutant mice. The specific aims of the Core are to perform modification of genes of interest in bacterial artificial chromosome (BAC) for use in transgenic mice; to isolate of primary tubule ceNs and conditionally immortalized cell lines from specific nephron segments of mutant mouse strains; to facilitate generation of conditional knockout and knockin gene targeting strategies and constructs and to provide general resources for investigators in mouse genetic applications. This Core will Coordinate with the Renal Physiology and Phenotyping Core to assist investigators couple mouse genetic services with physiological studies. It will coordinate with the Human Genetics and Clinical Research to integrate new human disease gene discoveries with animal and cell line models. There is inherent synergy in the mouse model and cell line components of this Core as it allowing investigators access to the spectrum from defined in vivo models to ex vivo cell line models with identical genetic makeup. The Core now adds a novel mechanism for cell line generation based on technology developed by Core faculty, an improved BAC recombineering methodology and a new service in conditional knockout or knockin allele generation based the BAC technology.