High-Throughput CRISPR Screening Identifies Genes Involved in Macrophage Viability and Inflammatory Pathways.
High-Throughput CRISPR Screening Identifies Genes Involved in Macrophage Viability and Inflammatory Pathways.
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DOI：
10.1016/j.celrep.2020.108541
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发表时间：
2020-12-29
期刊：
影响因子：
8.8
通讯作者：
Carpenter S
中科院分区：
文献类型：
--
作者：
Covarrubias S;Vollmers AC;Capili A;Boettcher M;Shulkin A;Correa MR;Halasz H;Robinson EK;O'Briain L;Vollmers C;Blau J;Katzman S;McManus MT;Carpenter S
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. Covarrubias et al. screen ~21,000 targets, generating a resource guide of genes required for macrophage viability as well as previously unidentified positive and negative regulators of NF-κB signaling. They identify regulatory elements within essential genes and show that membrane-bound TNF primarily functions in macrophages in an autocrine fashion to negatively regulate inflammation.
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影响因子：
39.3
作者：
Liu T;Zhang L;Joo D;Sun SC
通讯作者：
Sun SC
影响因子：
7.3
作者：
Bhatt D;Ghosh S
通讯作者：
Ghosh S
影响因子：
11.2
作者：
Ardestani S;Li B;Deskins DL;Wu H;Massion PP;Young PP
通讯作者：
Young PP
DOI：
10.1073/pnas.1307002110
发表时间：
2013-06-18
影响因子：
11.1
作者：
Kampmann, Martin;Bassik, Michael C.;Weissman, Jonathan S.
通讯作者：
Weissman, Jonathan S.
DOI：
10.1126/science.aat5011
发表时间：
2018-08-31
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Knott GJ;Doudna JA
通讯作者：
Doudna JA