Mar 31, 2020

CiBER-seq dissects genetic networks by quantitative CRISPRi profiling of expression phenotypes

BioRxiv : the Preprint Server for Biology
Eugenio UrdapilletaAlessandro Treves

Abstract

To realize the promise of CRISPR/Cas9-based genetics, approaches are needed to quantify a specific, molecular phenotype across genome-wide libraries of genetic perturbations. We address this challenge by profiling transcriptional, translational, and post-translational reporters using CRISPR interference with barcoded expression reporter sequencing (CiBER-seq). Our barcoding approach connects an entire library of guides to their individual phenotypic consequences using pooled sequencing. We show that CiBER-seq profiling fully recapitulates the integrated stress response (ISR) pathway in yeast. Genetic perturbations causing uncharged tRNA accumulation activated ISR reporter transcription. Surprisingly, tRNA insufficiency also activated the reporter, independent of the Gcn2 kinase that senses uncharged tRNAs. By uncovering alternate triggers for ISR activation, we illustrate how precise, comprehensive CiBER-seq profiling provides a powerful and broadly applicable tool for dissecting genetic networks.

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Mentioned in this Paper

Embryo
Entire Embryo
Neurons
Environment
Anatomical Space Structure
Caged molecule
Grid Cells
Neuronal
Rodent
5-methylchrysene

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