Vanderbilt O'Brien Kidney Center - Core C Cell and Genome Engineering

摘要 细胞和基因组工程的核心结构，以补充小鼠和人类的体内研究 通过在体内和体外对细胞进行遗传操作以及分离和繁殖特异性的 肾细胞这些服务将由一个工作人员提供，这是独特的经验， 方法论。 该中心将提供以下服务： 1)为肾脏病学界提供基因组工程方面的专业知识，特别是 关于提供CRISPR/Cas9和转座子载体以及将其用于 培养的肾细胞的稳定遗传修饰。 2)从小鼠中分离离散的细胞群（例如足细胞、近端小管细胞）， 作为原代细胞或永生化细胞培养以备将来使用。此外，它将分离蛋白质和/或 来自受损小鼠肾脏细胞特异性成分的mRNA用于深入分析。 3)从正常和患病的人肾脏分离足细胞和近端小管细胞， 进行转录组学和蛋白质组学分析。患者临床数据将链接到 通过电子记录的样本。

Abstract The cell and genome engineering core is structured to complement mouse and human in vivo studies by genetically manipulating cells both in vivo and in vitro as well as isolating and propagating specific renal cells. These services will be provided by a staff that is uniquely experienced with these methodologies. The Core will provide the following services: 1) Provide expertise with regards to genome engineering to the Nephrology community, specifically with respect to providing CRISPR/Cas9 and transposon vectors and expertise in using them for stable genetic modification of kidney cells in culture. 2) Isolate discrete cell populations (e.g. podocytes, proximal tubule cells) from mice that can be cultured as either primary or immortalized cells for future use. In addition, it will isolate protein and/or mRNA from cell-specific components of injured mouse kidneys for in depth analysis. 3) Isolate podocytes and proximal tubule cells from normal and diseased human kidneys that will be processed for transcriptomic and proteomic analyses. The patient clinical data will be linked to the samples via electronic records.