Perturb-seq: Single-cell CRISPRi screens interrogate the Unfolded Protein Response
Britt Adamson, PhD
Wednesday, November 29, 2017
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Chapters
Introduction
The Unfolded Protein Response is a branched stress response network with diverse inputs
Traditional approaches for evaluating transcriptional programs require a complexity tradeoff
Can we build a screening platform that measures complex phenotypes at genome-scale?
Pooled CRISPRi screening by single-cell RNA-seq: Perturb-seq
The Perturb-seq platform
Challenge 1: How to capture perturbation identity within single-cell transcriptomes?
sgRNA identity mapping is robust
What can Perturb-seq can do?
Clustering genes by correlation of bulk Perturb-seq expression profiles reveals functional relationships
What can Perturb-seq can do?
Systematic decoupling of transcriptional circuits: Perturb-seq after repression of three UPR arms
Decoupling of transcriptional programs with single-cell data
Decoupling transcriptional programs within Perturb-seq data from UPR-inducing sublibrary
What can Perturb-seq can do?
Do individual cells execute a heterogeneous UPR in response to homogeneous perturbation?
Bifurcated UPR induction in response to BiP (HSPA5) depletion
What can Perturb-seq can do?
Acknowledgements
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