Mouse Genetics and Cell Line Core

项目总结(见说明)： 在过去的十年里，在将技术应用于生物医学科学方面取得了巨大的进步。其中更显着的成就是全基因组测序的出现，使人们能够完整地了解可感染生物的基因组DNA结构，以及相关的分子生物学进展，使得能够对这些新发现的基因组序列进行全面操作。随之而来的是将肾脏疾病和功能的科学研究提升到哺乳动物肾脏体内精细化系统的能力。对肾脏疾病基本科学发现的验证现在取决于对转基因小鼠模型和自然产生的人类样本的研究。耶鲁大学乔治·M·奥布莱恩肾脏中心的小鼠遗传学和细胞系核心中心的首要目标是减少障碍，促进体内基于老鼠的技术在肾脏疾病研究中的应用，并促进将研究扩展到来自工程突变小鼠的体外基于细胞的系统。该核心的具体目标是对用于转基因小鼠的细菌人工染色体(BAC)中的感兴趣基因进行修改；从突变小鼠品系的特定肾单位片段中分离初级小管中心和条件永生化细胞系；促进条件敲除和敲除基因靶向策略和构建的生成，并为小鼠遗传学应用中的研究人员提供通用资源。该核心将与肾脏生理学和表型核心协调，以协助调查人员将老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。这一核心的小鼠模型和细胞系组件具有内在的协同作用，因为它使研究人员能够获得从定义的体内模型到具有相同基因组成的体外细胞系模型的光谱。Core现在增加了一种新的细胞系生成机制，该机制基于Core Facters开发的技术，改进的BAC重组工程方法，以及基于BAC技术的条件敲除或敲击等位基因生成的新服务。

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.