Functional Genomics

摘要-功能基因组学共享资源 由癌症中心管理的功能基因组学共享资源的中心目标是 为中心成员提供服务和工具，以在培养的细胞中进行RNAi和CRISPR-CAS9基因筛选。 细胞核心工作人员在整个项目过程中与中心教师互动，从项目开始， 概念化和可行性评估，直至对屏幕中确定的目标进行最终验证。 核心利用了Prebys中心和化学中心大量的HTS设备， 文库筛选核心。除了工作人员带来的丰富的专业知识外，核心还作为一个储存库， 用于与癌症中心研究人员共享的试剂、测定和技术信息。核心 提供多种用于功能基因组学分析的文库，包括全基因组siRNA文库，和 全面的miRNA激活剂和抑制剂文库，广泛的shRNA文库，以及靶向阵列和 全基因组合并的CRISPR文库。核心的主要工作是致力于通过RNAi进行遗传筛选， CRISPR-Cas9，尽管目前也有大量时间致力于细胞系的工程化。在核心内部， 主要实验室专注于细胞遗传学，而病毒载体实验室则专注于高质量载体 在一个有专家工作人员的集中设施中制备，并产生一系列复杂的病毒系统， 使核酸能够递送。这两个实验室还积极参与开发针对 研究人员的特殊需求。功能基因组学与其他共享的 SBP的资源。因此，Core利用了来自邻近的自动化和高通量成像支持， 化学文库筛选共享资源;与生物信息学共享资源交互以执行 原始数据的统计分析和设计定制CRISPR文库;并使用 基因组学共享资源来执行CRISPR-Cas9筛选。总的来说，功能基因组学核心 努力保持其在细胞遗传学和核酸输送的前沿地位， 最先进的技术，实施方法，以填补技术差距，并调整方法，以实现具体的 癌症中心的需求。在过去5年中，共有45个中心成员使用了该核心， 所有三个项目，它支持癌症中心成员至少29个癌症相关的出版物。

ABSTRACT – FUNCTIONAL GENOMICS SHARED RESOURCE The central goal of the Functional Genomics Shared Resource, which is managed by the Cancer Center, is to provide services and tools for Center members to perform RNAi and CRISPR-CAS9 genetic screens in cultured cells. Core staff interact with Center faculty members throughout their projects, starting with project conceptualization and feasibility assessment through to the final verification of targets identified in the screens. The Core utilizes the extensive amount of HTS equipment available in the Prebys Center and the Chemical Library Screening Core. In addition to the wealth of expertise brought by the staff, the Core serves as a repository for reagents, assays, and technical information, which are shared with Cancer Center researchers. The Core provides a variety of libraries for functional genomics analysis, including a genome-wide siRNA library, and comprehensive miRNA activator and inhibitor libraries, broad shRNA libraries, as well as targeted arrayed and genome-wide pooled CRISPR libraries. The Core's primary effort is dedicated to genetic screening via RNAi and CRISPR-Cas9, although significant time is also currently dedicated to engineering of cell lines. Within the Core, the main laboratory focuses on cellular genetics, while the viral vector laboratory specializes in high-quality vector preparation in a centralized facility with expert staff, and produces an array of sophisticated viral systems to enable nucleic acid delivery. Both laboratories are also actively involved in developing tools tailored to the specific needs of Center researchers. Functional Genomics operates in close connection with other Shared Resources at SBP. Thus, the Core utilizes automation and high-throughput imaging support from the adjacent Chemical Library Screening Shared Resource; interacts with the Bioinformatics Shared Resource to perform statistical analysis of the raw data and design custom CRISPR libraries; and uses the sequencing services of the Genomics Shared Resource to execute CRISPR-Cas9 screens. Overall, the Functional Genomics Core has strived to maintain its place at the forefront of cellular genetics and nucleic acid delivery by adopting state-of- the-art technologies, implementing methods to fill technology gaps, and adapting methods to fulfill the specific needs of the Cancer Center. In the past 5 years, the Core was used by a total of 45 Center members representing all three programs, and it supported at least 29 cancer-related publications by Cancer Center members.