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Chemical rescue of cleft palate and midline defects in conditional GSK-3β mice

Nature volume 446, pages 7982 (01 March 2007) | Download Citation

Abstract

Glycogen synthase kinase-3β (GSK-3β) has integral roles in a variety of biological processes, including development, diabetes, and the progression of Alzheimer’s disease1,2,3,4. As such, a thorough understanding of GSK-3β function will have a broad impact on human biology and therapeutics. Because GSK-3β interacts with many different pathways, its specific developmental roles remain unclear5. We have discovered a genetic requirement for GSK-3β in midline development. Homozygous null mice display cleft palate, incomplete fusion of the ribs at the midline and bifid sternum as well as delayed sternal ossification. Using a chemically regulated allele of GSK-3β (ref. 6), we have defined requirements for GSK-3β activity during discrete temporal windows in palatogenesis and skeletogenesis. The rapamycin-dependent allele of GSK-3β produces GSK-3β fused to a tag, FRB* (FKBP/rapamycin binding), resulting in a rapidly destabilized chimaeric protein. In the absence of drug, GSK-3βFRB*/FRB* mutants appear phenotypically identical to GSK-3β-/- mutants. In the presence of drug, GSK-3βFRB* is rapidly stabilized, restoring protein levels and activity6. Using this system, mutant phenotypes were rescued by restoring endogenous GSK-3β activity during two distinct periods in gestation. This technology provides a powerful tool for defining windows of protein function during development.

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Acknowledgements

We thank M. S. Dionne for critical reading of the manuscript; M. S. Dionne, M. M. Winslow, J. E. Gestwicki, J. H. Bayle, S. C. Kao and members of the Longaker and Crabtree laboratories for invaluable discussions; and J. Woodgett for the gift of GSK-3β knockout mice. K.J.L and M.T.L. are supported by the NIH, M.T.L. is also supported by the Oak Foundation and J.R.A. is a fellow of the Berry Foundation.

Author information

Author notes

    • Karen J. Liu
    •  & Joseph R. Arron

    Present addresses: Department of Craniofacial Development, King’s College London, London SE1 9RT, UK (K.J.L.); Department of Immunology Diagnostics, Genentech, 1 DNA Way, South San Francisco, California 94080, USA (J.R.A.)

    • Karen J. Liu
    •  & Joseph R. Arron

    These authors contributed equally to this work.

Affiliations

  1. The Department of Pathology,

    • Karen J. Liu
    • , Joseph R. Arron
    •  & Gerald R. Crabtree
  2. The Stanford Institute for Stem Cell Biology and Regenerative Medicine,

    • Karen J. Liu
    •  & Michael T. Longaker
  3. Department of Developmental Biology,

    • Kryn Stankunas
    •  & Gerald R. Crabtree
  4. Howard Hughes Medical Institute, and the

    • Gerald R. Crabtree
  5. Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Palo Alto, California 94305, USA.

    • Michael T. Longaker

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Competing interests

Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Corresponding authors

Correspondence to Karen J. Liu or Michael T. Longaker.

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    Supplementary Information

    This file contains Supplementary Methods and Supplementary Figures S1-S2 with legends.

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DOI

https://doi.org/10.1038/nature05557

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