期刊
CELL
卷 167, 期 7, 页码 1867-+出版社
CELL PRESS
DOI: 10.1016/j.cell.2016.11.048
关键词
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资金
- National Institutes of Health [P50 GM102706, U01 CA168370, R01 DA036858, R01 GM102790, R35 GM118061, F32 GM116331]
- NIH/NCI Pathway to Independence Award [K99 CA204602]
- European Commission [GA-2013-609409, EMBO ALTF 1193-2015]
- NDSEG Fellowship
- Klarman Cell Observatory
- NHGRI [P50 HG006193]
- HHMI
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.
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