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Nat Genet. 2018 Mar;50(3):460-471. doi: 10.1038/s41588-018-0054-7. Epub 2018 Feb 19.

A CRISPR-based screen for Hedgehog signaling provides insights into ciliary function and ciliopathies.

Author information

1
Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, USA. david.breslow@yale.edu.
2
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA. david.breslow@yale.edu.
3
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA.
4
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA.
5
Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
6
Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, USA.
7
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA. jameschen@stanford.edu.
8
Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, USA. jameschen@stanford.edu.
9
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA. maxence.nachury@ucsf.edu.
10
Department of Ophthalmology, UCSF, San Francisco, CA, USA. maxence.nachury@ucsf.edu.

Abstract

Primary cilia organize Hedgehog signaling and shape embryonic development, and their dysregulation is the unifying cause of ciliopathies. We conducted a functional genomic screen for Hedgehog signaling by engineering antibiotic-based selection of Hedgehog-responsive cells and applying genome-wide CRISPR-mediated gene disruption. The screen can robustly identify factors required for ciliary signaling with few false positives or false negatives. Characterization of hit genes uncovered novel components of several ciliary structures, including a protein complex that contains δ-tubulin and ε-tubulin and is required for centriole maintenance. The screen also provides an unbiased tool for classifying ciliopathies and showed that many congenital heart disorders are caused by loss of ciliary signaling. Collectively, our study enables a systematic analysis of ciliary function and of ciliopathies, and also defines a versatile platform for dissecting signaling pathways through CRISPR-based screening.

PMID:
29459677
PMCID:
PMC5862771
DOI:
10.1038/s41588-018-0054-7
[Indexed for MEDLINE]
Free PMC Article

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