CRISPR-Image Core

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.