CRISPR-Image Core

项目摘要-核心C CRISPR-图像 CRISPR技术正在给生物科学带来革命性的变化，并加速其发现。它们实现了高效的 哺乳动物细胞中基因的编辑、插入和失活。同样，高分辨率和高吞吐量 光学成像可以快速分析细胞和分子表型。这一核心将利用我们最近的 CRISPR和光学成像能力的进展以及我们在荧光方面的丰富经验 蛋白质分子生物学，创造了一个强大的新资源，体现在CRISPR-IMAGE核心中。 在这里，我们建议将现有资源与新资源结合起来，为科布雷提供所需的能力 (BioSNTR-II)。CRISPR-IMAGE核心将在 BioSNTR-II的研究项目。具体地说，这一核心将使在 在所有三个研究项目中都提出了细胞系和原代细胞。CRISPR-IMAGE核心有三个 特异性目的：1)CRISPR介导的基因破坏和荧光蛋白嵌合体在免疫和 炎症细胞；2)正常细胞或CRISPR编辑细胞中细胞结构的高分辨率成像；以及3) 细胞功能的高含量显微镜、流式细胞仪分析和全基因组CRISPR筛查 在经过基因编辑的细胞中。这个核心特别重要，因为它带来了尖端的分子生物学和 向研究项目提供显微技术，帮助他们在生物医学研究中产生强大的影响 并在国内和国际融资领域展开竞争。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，从而将基因组与物候组联系起来 进入压缩的工作流程。

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.