Vector and Transgenic Mouse Core

载体和转基因小鼠核心的总体目标是为糖尿病研究中心提供 华盛顿大学和大西雅图地区的附属研究人员， 在一些实施方案中，所述方法是过度表达、敲低、敲除、编辑或以其他方式改变RNA和蛋白质表达所必需的。 对培养细胞、分离组织和动物感兴趣。自上一次世界大战以来， 竞争性更新。新服务提供CRISPR（定期间隔的短回文聚类） 已经添加了用于鉴定编辑的基因的载体、试剂和方法，并且已经添加了用于鉴定编辑的基因的Cas9载体、试剂和方法的数量。 可用的病毒载体血清型已大大扩展。这些新服务的使用率很高，现在 提供了核心的大部分工作。相反，那些不那么先进的服务， 经常被要求删除。核心的具体目标是提供以下服务， 附属研究者：（1）生产腺相关病毒载体（AAV）、慢病毒载体、泡沫病毒 载体和γ逆转录病毒载体用于动物、组织和细胞;（2）基因组编辑（编辑，敲除， 通过CRISPR/Cas9在细胞和小鼠中敲除（敲入）;（3）具有成本效益的基因工程小鼠生产 通过华盛顿大学的临床前研究和 （四）利用分子生物学技术筛选和分析基因编辑的效果; （5）咨询和培训。在本供资期内，核心方案的生产力很高， 增加了新的服务，预计将显着提高生产力和可用性的糖尿病研究 中心附属研究人员，以满足该中心的目标，加强对糖尿病，肥胖和相关的研究 大西雅图地区及其他地区的疾病。

The overall objective of the Vector and Transgenic Mouse Core is to provide Diabetes Research Center affiliate investigators at the University of Washington and the Greater Seattle area with state-of-the-art vectors necessary to overexpress, knockdown, knockout, edit, or otherwise alter expression of RNAs and proteins of interest in cultured cells, isolated tissues, and animals. The Core has considerably evolved since the last competitive renewal. New services provide CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 vectors, reagents and methods to identify edited genes have been added, and the number of viral vector serotypes available has been greatly expanded. These newer services are highly used, and now provide the bulk of the Core's work. Conversely, services that are less cutting-edge and are no longer frequently requested have been removed. The specific aims of the Core are to provide the following services to affiliate investigators: (1) Production of adeno-associated viral vectors (AAVs), lentiviral vectors, foamy virus vectors, and gammaretroviral vectors for use in animals, tissues and cells; (2) Genome editing (editing, knock- out, knock-in) in cells and mice by CRISPR/Cas9; (3) Cost-effective production of genetically engineered mice by CRISPR/Cas9 and cryopreservation through the University of Washington Preclinical Research and Transgenic Services; (4) Screening and analysis of effects of gene editing by molecular biology techniques; and (5) Consultation and training. The Core has been highly productive in the current funding period and has added new services that are expected to significantly increase productivity and usability for Diabetes Research Center affiliate investigators to meet the Center's goal to enhance research in diabetes, obesity and related disorders in the Greater Seattle area and beyond.