ReporterSeq Reveals Genome-wide Dynamic Modulators of the Heat Shock Response Across Diverse Stressors

Overview

Journal Elife

Specialty Biology

Date 2021 Jul 5

PMID 34223816

Citations 13

Authors

Brian D Alford

Eduardo Tassoni-Tsuchida

Danish Khan

Jeremy J Work

Gregory Valiant

Onn Brandman

Affiliations

Soon will be listed here.

Abstract

Understanding cellular stress response pathways is challenging because of the complexity of regulatory mechanisms and response dynamics, which can vary with both time and the type of stress. We developed a reverse genetic method called ReporterSeq to comprehensively identify genes regulating a stress-induced transcription factor under multiple conditions in a time-resolved manner. ReporterSeq links RNA-encoded barcode levels to pathway-specific output under genetic perturbations, allowing pooled pathway activity measurements via DNA sequencing alone and without cell enrichment or single-cell isolation. We used ReporterSeq to identify regulators of the heat shock response (HSR), a conserved, poorly understood transcriptional program that protects cells from proteotoxicity and is misregulated in disease. Genome-wide HSR regulation in budding yeast was assessed across 15 stress conditions, uncovering novel stress-specific, time-specific, and constitutive regulators. ReporterSeq can assess the genetic regulators of any transcriptional pathway with the scale of pooled genetic screens and the precision of pathway-specific readouts.

Citing Articles

Mapping the Genetic Architecture of the Adaptive Integrated Stress Response in .

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