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调查人员拥有最先进的媒介 过表达、敲除、敲除、编辑或以其他方式改变RNA和蛋白质的表达所必需的 对培养的细胞、分离的组织和动物感兴趣。核心自上一年以来已经有了相当大的演变 竞争性更新。新服务提供CRISPR(集群规则间隔短回文 Repeats)/Cas9载体、试剂和识别编辑基因的方法已被添加，并且 现有的病毒载体血清型已大大扩大。这些较新的服务使用率很高，现在 提供核心的大部分工作。相反，不那么尖端的服务不再是 经常被请求的内容已被删除。核心的具体目标是提供以下服务 附属调查：(1)腺相关病毒载体(AAVs)、慢病毒载体、泡沫病毒的生产 用于动物、组织和细胞的载体和伽马逆转录病毒载体；(2)基因组编辑(编辑、敲击- 通过CRISPR/Cas9导入细胞和小鼠；(3)低成本生产基因工程小鼠 由CRISPR/Cas9和冷冻保存通过华盛顿大学临床前研究和 转基因服务；(4)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。核心基金在本供资期间卓有成效，并已 添加了新服务，有望显著提高糖尿病研究的工作效率和可用性 中心附属调查人员满足中心加强糖尿病、肥胖症及相关研究的目标 大西雅图地区及更远地区的混乱。

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.