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CRISPR Screens Uncover Genes that Regulate Target Cell Sensitivity to the Morphogen Sonic Hedgehog.

Developmental cell | 2018

To uncover regulatory mechanisms in Hedgehog (Hh) signaling, we conducted genome-wide screens to identify positive and negative pathway components and validated top hits using multiple signaling and differentiation assays in two different cell types. Most positive regulators identified in our screens, including Rab34, Pdcl, and Tubd1, were involved in ciliary functions, confirming the central role for primary cilia in Hh signaling. Negative regulators identified included Megf8, Mgrn1, and an unannotated gene encoding a tetraspan protein we named Atthog. The function of these negative regulators converged on Smoothened (SMO), an oncoprotein that transduces the Hh signal across the membrane. In the absence of Atthog, SMO was stabilized at the cell surface and concentrated in the ciliary membrane, boosting cell sensitivity to the ligand Sonic Hedgehog (SHH) and consequently altering SHH-guided neural cell-fate decisions. Thus, we uncovered genes that modify the interpretation of morphogen signals by regulating protein-trafficking events in target cells.

Pubmed ID: 29290584 RIS Download

Antibodies used in this publication

Associated grants

  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM112988
  • Agency: Cancer Research UK, United Kingdom
    Id: FC001051
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM118082
  • Agency: Medical Research Council, United Kingdom
  • Agency: Wellcome Trust, United Kingdom
    Id: WT098326MA
  • Agency: Wellcome Trust, United Kingdom
  • Agency: Arthritis Research UK, United Kingdom
    Id: FC001051
  • Agency: NIGMS NIH HHS, United States
    Id: DP2 GM105448
  • Agency: Wellcome Trust, United Kingdom
    Id: FC001051

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