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 - 核心C CRISPR图像 CRISPR技术正在革新生物科学中的发现。它们提高了高效基因在哺乳动物细胞中的编辑，插入和失活。同样，高分辨率和高通量光学成像可以快速分析细胞和分子表型。这个核心将利用我们的最新 CRISPR和光学成像功能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，创建一个强大的新资源，体现在CRISPR形象核心中。在这里，我们建议将现有资源与新资源团结起来，以提供此毛绒所需的功能（biosntr-ii）。 CRISPR-图像核心将在该核心中实现尖端的基因型对表型分析 Biosntr-II的研究项目。具体而言，该核心将使目标基因敲除在所有三个研究项目中都提出了细胞系和主要细胞。 CRISPR形象核心有三个具体目的：1）在免疫和炎性细胞； 2）正常或CRISPR编辑的细胞中细胞结构的高分辨率成像； 3）高素质显微镜，流式细胞仪分析和全基因组CRISPR筛选细胞功能在基因编辑的细胞中。该核心尤其重要，因为它带来了领先的分子生物学和研究项目的显微镜技术，以帮助他们对生物医学研究产生有力的影响并参加国家和国际资助领域。这个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，从而将基因组与酚类联系起来进入压缩工作流程。