CRISPR-Image Core

项目概要-核心C CRISPR-IMAGE CRISPR技术正在彻底改变和加速生物科学的发现。使高效 编辑、插入和失活哺乳动物细胞中的基因。同样，高分辨率和高通量 光学成像能够快速分析细胞和分子表型。这个核心将利用我们最近的 CRISPR和光学成像能力的进步，以及我们在荧光领域的丰富经验，沿着 蛋白质分子生物学，创造一个强大的新资源，体现在CRISPR-IMAGE核心。 在这里，我们建议将现有资源与新资源结合起来，为COBRE提供所需的能力 （BioSNTR-II）。CRISPR-IMAGE Core将在生物学领域内实现尖端的基因型到表型分析。 BioSNTR-II的研究项目。具体地说，这个核心将能够在细胞中产生靶向基因敲除， 细胞系和原代细胞在所有三个研究项目中提出。CRISPR-IMAGE核心有三个 具体目的：1）CRISPR介导的基因破坏和荧光蛋白嵌合体在免疫和 2）正常或CRISPR编辑的细胞中细胞结构的高分辨率成像;和 高内涵显微镜、流式细胞术分析和细胞功能的全基因组CRISPR筛选 在基因编辑的细胞中。这个核心特别重要，因为它带来了前沿的分子生物学， 显微镜技术的研究项目，以帮助他们在生物医学研究产生强大的影响 并在国内和国际融资舞台上竞争。这个核心是高度创新的，因为它直接 将CRISPR基因编辑与光学显微镜方法相结合， 进入压缩工作流程。

Project Summary – Core C CRISPR-IMAGE CRISPR technologies are revolutionizing and accelerating discovery in bioscience. They enable efficient editing, insertion and deactivation of genes in mammalian cells. Likewise, high-resolution and high-throughput optical imaging enables rapid analysis of cellular and molecular phenotypes. This core will leverage our recent advances in CRISPR and optical imaging capabilities along with our extensive experience in fluorescent protein molecular biology, to create a powerful new resource, embodied within the CRISPR-IMAGE Core. Here, we propose to unite existing resources with new ones to provide the needed capabilities for this COBRE (BioSNTR-II). The CRISPR-IMAGE Core will enable cutting-edge genotype-to-phenotype analysis within the research projects of BioSNTR-II. Specifically, this core will enable creation of the targeted gene knockouts in cell lines and primary cells proposed in all three of the research projects. The CRISPR-IMAGE Core has three specific aims: 1) CRISPR-mediated gene disruption and fluorescent protein chimera expression in immune and inflammatory cells; 2) High-resolution imaging of cellular structures in normal or CRISPR-edited cells; and 3) High-content microscopy, flow cytometric analysis, and whole-genome CRISPR screening of cellular functions in gene-edited cells. This core is particularly significant because it brings leading-edge molecular biology and microscopy technologies to the research projects to help them make potent impacts in biomedical research and to compete in the national and international funding arenas. This core is highly innovative in that it directly combines CRISPR gene-editing with optical microscopy methods thereby linking genome-to-phenome approaches into a compressive workflow.