2. Academic Validation
  3. A Genome-wide CRISPR Death Screen Identifies Genes Essential for Oxidative Phosphorylation

A Genome-wide CRISPR Death Screen Identifies Genes Essential for Oxidative Phosphorylation

  • Cell Metab. 2016 Dec 13;24(6):875-885. doi: 10.1016/j.cmet.2016.08.017.
Jason D Arroyo 1,  Alexis A Jourdain 1,  Sarah E Calvo 1,  Carmine A Ballarano 1,  John G Doench 2,  David E Root 2,  Vamsi K Mootha 3
Affiliations

Affiliations

  • 1 Department of Molecular Biology and Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
  • 2 Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
  • 3 Department of Molecular Biology and Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: vamsi@hms.harvard.edu.
PMID: 27667664 DOI: 10.1016/j.cmet.2016.08.017
Abstract

Oxidative Phosphorylation (OXPHOS) is the major pathway for ATP production in humans. Deficiencies in OXPHOS can arise from mutations in either mitochondrial or nuclear genomes and comprise the largest collection of inborn errors of metabolism. At present we lack a complete catalog of human genes and pathways essential for OXPHOS. Here we introduce a genome-wide CRISPR "death screen" that actively selects dying cells to reveal human genes required for OXPHOS, inspired by the classic observation that human cells deficient in OXPHOS survive in glucose but die in galactose. We report 191 high-confidence hits essential for OXPHOS, including 72 underlying known OXPHOS diseases. Our screen reveals a functional module consisting of NGRN, WBSCR16, RPUSD3, RPUSD4, TRUB2, and FASTKD2 that regulates the mitochondrial 16S rRNA and intra-mitochondrial translation. Our work yields a rich catalog of genes required for OXPHOS and, more generally, demonstrates the power of death screening for functional genomic analysis.

Keywords

16S RNA; CRISPR; Mitochondria; OXPHOS; death screen; mitochondrial ribosome; neugrin; pseudouridine; pseudouridylation; synthetic lethality.

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