Cell and Animal Model Core

在我们的研究计划中，主要从p53调节器和效应器的体外模型中获得的见解导致了对体内验证和使用体内模型进一步了解的能力的迫切需要。在P53范例中正在积极研究的表型不再是简单的细胞死亡、生长停滞或衰老的问题。相反，活体内P53激活的作用可能会在不同的细胞和组织类型中产生不同的后果，并且可能会随着发育阶段的不同而不同。细胞和动物模型核心(核心B)将衍生和维持人类和小鼠细胞系，以及项目使用的早期传代等份冷冻原代细胞的充足库存。它将作为单抗和经过验证的RNAi试剂的储存库。Core还将获得并维护稳定使用的小鼠模型(转基因、基因敲除等)。以及从这些小鼠中提取的原代细胞，用于分配给该计划的研究人员。我们的生产性和协作性项目的研究工作极大地受益于体内模型组件，该组件保持了小型繁殖 我们的研究人员为进行遗传和细胞生物学研究而急需的小鼠品系的群体。通过为小鼠育种和遗传分析以及共享免疫组织化学提供共同的技术支持，屏障设施中动物研究的高昂成本可以大大降低。几个研究人员和该计划内的项目共享使用相同的转基因菌株，进一步强调了成本效益。最后，该核心将为基因表达分析(蛋白质和信使核糖核酸原位)准备组织和/或胚胎提供专门知识和指导。

Insights gained primarily from in vitro models of p53 regulators and effectors within our research program have led to a critical need for in vivo validation and the ability to gain further understanding using in vivo models. The phenotypes that are actively being examined in the p53 paradigm are no longer simply a matter of cell death, growth arrest or senescence. Instead, the role of p53 activation in vivo will likely have different consequences In different cell and tissue types and will likely differ with regard to developmental stages. The Cell and Animal Model Core (Core B) will derive and maintain human and mouse cell lines as well as adequate stocks of early passage aliquots of frozen primary cells utilized by the projects. It will serve as a repository for monoclonal antibodies and validated RNAi reagents. The Core will also acquire and maintain steadily used mouse models (transgenic, knockouts, etc.) and primary cells derived from these mice for distribution to the investigators in the Program. The research efforts of our productive and collaborative program greatly benefit from an in vivo models component, which maintains small breeding colonies of critically needed mouse strains for genetic and cell biologic investigations by our investigators. The high cost of animal studies in barrier facilities can be reduced substantially by the availability of shared technical support for mouse breeding and genetic analyses and for shared immunohistochemistry. The cost effectiveness is further emphasized by the shared use of the same genetically modified strains by several investigators and projects within the Program. Lastly, the core will provide expertise and guidance in the preparation of tissue and/or embryos for analyses of gene expression (protein and mRNA in situ).