A Multiplexed Single-Cell CRISPR Screening Platform Enables Systematic Dissection of the Unfolded Protein Response

被引：674

作者：

Adamson, Britt ^{[1
,2
,3
,4
]}

Norman, Thomas M. ^{[1
,2
,3
,4
]}

Jost, Marco ^{[1
,2
,3
,4
,5
]}

Cho, Min Y. ^{[1
,2
,3
,4
]}

Nunez, James K. ^{[1
,2
,3
,4
]}

Chen, Yuwen ^{[1
,2
,3
,4
]}

Villalta, Jacqueline E. ^{[1
,2
,3
,4
]}

Gilbert, Luke A. ^{[1
,2
,3
,4
]}

Horlbeck, Max A. ^{[1
,2
,3
,4
]}

Hein, Marco Y. ^{[1
,2
,3
,4
]}

Pak, Ryan A. ^{[1
,8
]}

Gray, Andrew N. ^{[5
,13
]}

Gross, Carol A. ^{[5
,6
,7
]}

Dixit, Atray ^{[9
,10
]}

Parnas, Oren ^{[10
,12
]}

Regev, Aviv ^{[10
,11
]}

Weissman, Jonathan S. ^{[1
,2
,3
,4
]}

机构：

[1] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94158 USA

[2] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94158 USA

[3] Univ Calif San Francisco, Calif Inst Quantitat Biomed Res, San Francisco, CA 94158 USA

[4] Univ Calif San Francisco, Ctr RNA Syst Biol, San Francisco, CA 94158 USA

[5] Univ Calif San Francisco, Dept Microbiol & Immunol, San Francisco, CA 94158 USA

[6] Univ Calif San Francisco, Dept Cell & Tissue Biol, San Francisco, CA 94158 USA

[7] Univ Calif San Francisco, Integrat Program Quantitat Biol, San Francisco, CA 94158 USA

[8] Univ Calif Berkeley, Innovat Genom Initiat, Berkeley, CA 94720 USA

[9] Harvard Mit Div Hlth Sci & Technol, Cambridge, MA 02142 USA

[10] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[11] MIT, Dept Biol, Howard Hughes Med Inst, Cambridge, MA 02140 USA

[12] Hebrew Univ Jerusalem, Hadassah Med Sch, Lautenberg Ctr Gen & Tumor Immunol, BioMed Res Inst Israel Canada,Fac Med IMRIC, IL-91120 Jerusalem, Israel

[13] New England Biolabs Inc, Ipswich, MA 01938 USA

来源：

CELL | 2016年 / 167卷 / 07期

基金：

美国国家卫生研究院;

关键词：

ER-STRESS; RNA; GENES; ACTIVATION; EXPRESSION; ATF6; PERK; CAS9; SEQ; DNA;

D O I：

10.1016/j.cell.2016.11.048

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Functional genomics efforts face tradeoffs between number of perturbations examined and complexity of phenotypes measured. We bridge this gap with Perturb-seq, which combines droplet-based singlecell RNA-seq with a strategy for barcoding CRISPRmediated perturbations, allowing many perturbations to be profiled in pooled format. We applied Perturb-seq to dissect the mammalian unfolded protein response (UPR) using single and combinatorial CRISPR perturbations. Two genome-scale CRISPR interference (CRISPRi) screens identified genes whose repression perturbs ER homeostasis. Subjecting similar to 100 hits to Perturb-seq enabled highprecision functional clustering of genes. Single-cell analyses decoupled the three UPR branches, revealed bifurcated UPR branch activation among cells subject to thesameperturbation, and uncovered differential activation of the branches across hits, including an isolated feedback loop between the translocon and IRE1 alpha. These studies provide insight intohowthe three sensors ofERhomeostasis monitor distinct types of stress and highlight the ability of Perturb-seq to dissect complex cellular responses.

引用

页码：1867 / +

页数：37

共 44 条

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