Shared Resource-Transgenic Core

转基因核心共享资源-项目摘要/摘要 转基因核心的主要目标是为癌症中心成员和其他研究人员提供 索尔克研究所获得创建转基因小鼠模型的尖端技术。这些 转基因小鼠代表了关键的遗传工具，使研究人员能够模拟人类疾病， 分析和功能操纵特定基因，并测试潜在的治疗干预措施。实现 为了实现这一目标，转基因核心提供了最先进的基因组编辑服务，以及 产生和维持转基因小鼠。具体服务包括：1）将DNA构建体显微注射到 早期小鼠胚胎，2）CRISPR/Cas9显微注射，3）慢病毒显微注射，4）基因- 靶向小鼠胚胎干细胞（ES）进入囊胚，5）体外受精（IVF），6）胚胎和精子 冷冻保存，7）从非标准来源提供给研究所的小鼠品系的再衍生，8）基因- 小鼠ES细胞中的靶向，9）靶向载体设计和构建，10）ES细胞的从头衍生 系，11）ES细胞扩增和用于显微注射的制备，和12） DR 4和CF 1小鼠胚胎饲养细胞。此外，核心还提供注射人类 胚胎干细胞系和诱导多能干（iPS）细胞系植入免疫缺陷小鼠，以评估 形成畸胎瘤的能力最近，转基因核心已经扩大了服务范围，提供： 在修饰的ES细胞中确认基因靶向的服务，转基因小鼠的基于PCR的基因分型， 和支原体检测因此，转基因核心旨在促进癌症中心成员的研究， 产生转基因的癌症小鼠模型，冷冻保存目前没有的小鼠品系， 使用，重新衍生由非标准来源提供的小鼠品系，产生基于同源重组的 基因靶向小鼠ES细胞系，提供免费咨询，以匹配当前的转基因技术， 研究者的需要，并提供免费培训，在所有程序涉及生产转基因动物。

Transgenic Core Shared Resource - Project Summary/Abstract The primary goal of the Transgenic Core is to provide Cancer Center members and other investigators at the Salk Institute access to cutting-edge technologies for creating genetically altered mouse models. These transgenic lines of mice represent critical genetic tools that enable researchers to model human diseases, to analyze and functionally manipulate specific genes, and to test potential therapeutic interventions. To achieve this goal, the Transgenic Core provides state-of-the genome editing services, as well as services required to generate and maintain transgenic mice. Specific services include: 1) the microinjection of DNA constructs into early mouse embryos, 2) CRISPR/Cas9 microinjections, 3) lentiviral microinjections, 4) microinjection of gene- targeted mouse embryonic stem (ES) cells into blastocysts, 5) in-vitro fertilization (IVF), 6) embryo and sperm cryopreservation, 7) re-derivation of mouse lines provided to the Institute from non-standard sources, 8) gene- targeting in mouse ES cells, 9) targeting vector design and construction, 10) de novo derivation of ES cell lines, 11) ES cell expansion and preparation for microinjection, and 12) the generation and maintenance of DR4 and CF1 mouse embryonic feeder cells. In addition, the Core offers services for the injection of human embryonic stem cell lines and induced pluripotent stem (iPS) cell lines into immunodeficient mice to assess their ability to form teratomas. Recently the Transgenic Core has expanded services to offer: Southern blot services for confirmation of gene targeting in modified ES cells, PCR-based genotyping of transgenic mice, and mycoplasma testing. Thus, the Transgenic Core seeks to facilitate research of Cancer Center members by generating genetically modified mouse models of cancer, cryopreserving mouse lines that are not currently in use, re-deriving mouse lines provided by non-standard sources, generatimg homologous recombination-based gene targeted mouse ES cell lines, providing free consultations to match current transgenic technologies to investigator needs, and providing free training in all procedures involved in producing transgenic animals.