CRISPR-Image Core

CRISPR-图像核心

基本信息

批准号：
10593076
负责人：
Adam David Hoppe
金额：
$ 38.92万
依托单位：
SOUTH DAKOTA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-20 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10593076
关键词：
Acceleration Attention Biochemical Biological Assay Biological Sciences Biomedical Research CRISPR imaging CRISPR interference CRISPR screen CRISPR/Cas technology Cell Death Cell Line Cell Separation Cell Survival Cell physiology Cells Cellular Morphology Cellular Structures Centers of Research Excellence Chimeric Proteins Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Development Dose Ensure Evaluation Faculty Fee-for-Service Plans Flow Cytometry Fluorescence Fluorescence Recovery After Photobleaching Funding Gene Expression Generations Genes Genetic Genome Genomics Genotype Guide RNA Image Analysis Immune Inflammation Inflammatory International Knock-out Light Link Macrophage Mammalian Cell Mediating Methods Microscope Microscopy Modification Molecular Biology Oligonucleotides Open Reading Frames Optics Organelles Pathway Analysis Pathway interactions Pharmaceutical Preparations Phenotype Population Postdoctoral Fellow Principal Investigator Production Productivity Proteins Research Research Project Grants Research Support Resolution Resources Ribonucleoproteins Schools Services Signaling Protein South Dakota Speed Stimulus Students TNF gene Technology Time Training Training Support Translational Research analysis pipeline cytokine deep sequencing endonuclease experience fluorescence imaging gene complementation gene product graduate student high resolution imaging imaging capabilities industry partner innovation insertion/deletion mutation insight knockout gene lentivirally transduced live cell imaging member molecular phenotype novel optical imaging phenome programs rapid technique response screening tenure track three-dimensional visualization tool transcriptome whole genome

项目摘要

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.

项目摘要 – Core C CRISPR-IMAGE CRISPR 技术正在彻底改变并加速生物科学的发现，它们可以实现高效的发现。哺乳动物细胞中基因的编辑、插入和失活同样具有高分辨率和高通量。光学成像能够快速分析细胞和分子表型，该核心将利用我们最近的成果。 CRISPR 和光学成像能力的进步以及我们在荧光方面的丰富经验蛋白质分子生物学，创造强大的新资源，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 COBRE 提供所需的功能 (BioSNTR-II) CRISPR-IMAGE 核心将实现尖端的基因型到表型分析。具体来说，该核心将能够在 BioSNTR-II 的研究项目中创建目标基因敲除。所有三个研究项目中提出的细胞系和原代细胞都有三个。具体目标：1) CRISPR介导的基因破坏和荧光蛋白嵌合体在免疫和免疫中的表达炎症细胞；2) 正常细胞或 CRISPR 编辑细胞的高分辨率成像；高内涵显微镜、流式细胞术分析和细胞功能的全基因组 CRISPR 筛选在基因编辑细胞中，这一核心尤为重要，因为它带来了前沿的分子生物学和将显微镜技术应用到研究项目中，帮助他们对生物医学研究产生重大影响并在国内和国际融资领域竞争，这一核心具有高度创新性，因为它直接。将 CRISPR 基因编辑与光学显微镜方法相结合，从而将基因组与表型组联系起来进入压缩工作流程。