Core 2: In vivo cancer modeling and screening core facility

核心2：体内癌症建模和筛查核心设施

• 批准号: 9766838
• 负责人: Sidi Chen
• 金额: $ 27.24万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9766838
• 关键词: Animal Model; CRISPR library; CRISPR/Cas technology; Cancer Control; Cancer Model; Cell Culture Techniques; Cells; Collaborations; Communities; Complex; Core Facility; Custom; Disease model; Gene Mutation; General Population; Generations; Genes; Genetic; Genetic Models; Investigation; Lead; Libraries; Malignant Neoplasms; Mediating; Metastatic Neoplasm to the Lung; Modeling; Mus; Mutation; Neoplasm Transplantation; Phenotype; Population; Production; Proto-Oncogene Proteins c-akt; Research; Research Personnel; Research Project Grants; Research Support; Resources; Scientist; Students; System; Systems Analysis; Systems Biology; Technology; Training and Education; Viral; Viral Vector; Virus; base; cancer education; cell motility; design; gene discovery; genome-wide; high throughput screening; in vivo; in vivo evaluation; mammalian genome; mouse genome; mouse model; mutant; new therapeutic target; novel; particle; screening; somatic cell gene editing; tumor; vector; virtual animals

PROJECT SUMMARY/ABSTRACT: Core 2 We will establish a core facility for in vivo cancer modeling and screening. This core facility (core 2 in the cancer systems biology center) will consist of several sets of functional components, defined as modules, including vector design and genome-scale library construction, large-scale cell culture and virus production, tumor transplantation and in vivo pooled screening, CRISPR/Cas9 mediated somatic genome editing for invasive phenotype analysis. In preliminary studies, we have applied in vivo somatic genome editing to generate tumor models of specific driver genes in mice, and demonstrated in vivo screening in a lung metastasis animal model. We will build four modules in this core, including the following: Module 1. Facility for vector design and genome-scale library construction. This module supports functional investigations of the mammalian genome that can reveal how genetic alterations lead to changes in phenotype, for example cellular invasiveness phenotype described in the Projects 1 and 2. Module 2. Facility for large-scale cell culture, viral vector production and transduction. This will serve for two purposes: (1) generation of customized viral particles for in vivo studies involving animals models for Projects 1 and 2 as well as other collaborators; (2) high-throughput screening applications that requires transduction of a population of cells with highly complex libraries. Module 3. Facility for tumor transplantation and in vivo pooled screening. This module will setup a facility with two capacities: (1) tumor transplantation and (2) in vivo pooled screening. This will serve for the purpose of validating the genes discovered in the projects in mouse models, to discover new genes with invasive phenotypes using in vivo screens, or to enable collaborative research for discovery of new drug targets. Module 4. Facility for CRISPR/Cas9 mediated somatic genome editing, for somatic genome editing in various animals for virtually any loci in the mouse genome. We will establish this platform and utilize it for research described in the projects and for setting up collaborations with various Yale investigators. Specifically, we will utilize CRISPR library approach in conjunction with the Rapid Analysis of Cell migration Enhancement (RACE) system described in the Projects, to identify how genetic alterations lead to changes in cellular invasiveness phenotype. In addition, we will utilize these modules in the core to build novel models of WNK1, NKCC1 and their targets, as well as combinations of mutant ERK, AKT and downstream genes. In summary, this core will be tightly integrated into the U54 center, provide a powerful technology and resource platform for in vivo cancer systems biology, and support Research Projects in the center as well as other collaborators at Yale and the wider scientific community. Since there is no facility currently existing at Yale providing similar functions, this new core will facilitate the establishment of the first in vivo cancer modeling and screening core facility at Yale. We will also utilize this core facility for promoting education and training, serving as a base for education of cancer researchers, scientists, students, and the general public.

项目摘要/摘要：核心 2 我们将建立体内癌症建模和筛查的核心设施。该核心设施（核心 2 癌症系统生物学中心）将由几组功能组件组成，定义为模块， 包括载体设计和基因组规模的文库构建、大规模细胞培养和病毒生产， 肿瘤移植和体内混合筛选，CRISPR/Cas9介导的体细胞基因组编辑 侵袭性表型分析。在初步研究中，我们已将体内体细胞基因组编辑应用于 在小鼠中生成特定驱动基因的肿瘤模型，并在肺中进行体内筛选 转移动物模型。我们将在此核心构建四个模块，包括以下内容： 模块 1. 设施 用于载体设计和基因组规模的文库构建。该模块支持功能调查 例如，哺乳动物基因组可以揭示遗传改变如何导致表型变化 项目 1 和 2 中描述的细胞侵袭表型。模块 2.大规模细胞设施 培养、病毒载体生产和转导。这将有两个目的：（1）生成 用于涉及项目 1 和 2 以及其他动物模型的体内研究的定制病毒颗粒 合作者； (2) 需要转导细胞群的高通量筛选应用 高度复杂的库。模块 3. 肿瘤移植和体内混合筛选设施。这 该模块将建立一个具有两种能力的设施：（1）肿瘤移植和（2）体内汇总筛选。这 将用于在小鼠模型中验证项目中发现的基因，以发现新的基因 使用体内筛选具有侵入性表型的基因，或进行协作研究以发现新的 药物靶点。模块 4. 用于体细胞基因组的 CRISPR/Cas9 介导的体细胞基因组编辑设施 在各种动物中对小鼠基因组中的几乎任何位点进行编辑。我们将建立这个平台并利用 它用于项目中描述的研究以及与耶鲁大学的各个研究人员建立合作。 具体来说，我们将利用 CRISPR 文库方法结合细胞迁移快速分析 项目中描述的增强（RACE）系统，以确定基因改变如何导致变化 细胞侵袭表型。此外，我们将在核心中利用这些模块来构建新的模型 WNK1、NKCC1 及其靶标，以及突变 ERK、AKT 和下游基因的组合。在 综上所述，该核心将紧密集成到U54中心，提供强大的技术和资源 体内癌症系统生物学平台，并支持该中心以及其他研究项目 耶鲁大学和更广泛的科学界的合作者。由于耶鲁大学目前没有任何设施 该新核心提供类似的功能，将有助于建立第一个体内癌症模型和 耶鲁大学的筛选核心设施。我们还将利用这一核心设施来促进教育和培训，服务 作为癌症研究人员、科学家、学生和公众的教育基地。