清脆的
生物
效应器
功能(生物学)
基因
遗传筛选
单元格排序
巨噬细胞
免疫系统
细胞生物学
自分泌信号
计算生物学
遗传学
细胞
表型
细胞培养
体外
作者
Sergio Covarrubias,Apple Cortez Vollmers,Allyson Capili,Michael Boettcher,Aaron Shulkin,Michele Ramos Correa,Haley Halasz,Elektra K. Robinson,Laura O’Briain,Christopher Vollmers,James Blau,Sol Katzman,Michael T. McManus,Susan Carpenter
出处
期刊:Cell Reports
[Elsevier]
日期:2020-12-01
卷期号:33 (13): 108541-108541
被引量:29
标识
DOI:10.1016/j.celrep.2020.108541
摘要
Macrophages are critical effector cells of the immune system, and understanding genes involved in their viability and function is essential for gaining insights into immune system dysregulation during disease. We use a high-throughput, pooled-based CRISPR-Cas screening approach to identify essential genes required for macrophage viability. In addition, we target 3′ UTRs to gain insights into previously unidentified cis-regulatory regions that control these essential genes. Next, using our recently generated nuclear factor κB (NF-κB) reporter line, we perform a fluorescence-activated cell sorting (FACS)-based high-throughput genetic screen and discover a number of previously unidentified positive and negative regulators of the NF-κB pathway. We unravel complexities of the TNF signaling cascade, showing that it can function in an autocrine manner in macrophages to negatively regulate the pathway. Utilizing a single complex library design, we are capable of interrogating various aspects of macrophage biology, thus generating a resource for future studies.
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