Pierre Robin sequence (PRS) is an important subgroup of cleft palate. We report several lines of evidence for the existence of a 17q24 locus underlying PRS, including linkage analysis results, a clustering of translocation breakpoints 1.06–1.23 Mb upstream of SOX9, and microdeletions both 1.5 Mb centromeric and 1.5 Mb telomeric of SOX9. We have also identified a heterozygous point mutation in an evolutionarily conserved region of DNA with in vitro and in vivo features of a developmental enhancer. This enhancer is centromeric to the breakpoint cluster and maps within one of the microdeletion regions. The mutation abrogates the in vitro enhancer function and alters binding of the transcription factor MSX1 as compared to the wild-type sequence. In the developing mouse mandible, the 3-Mb region bounded by the microdeletions shows a regionally specific chromatin decompaction in cells expressing Sox9. Some cases of PRS may thus result from developmental misexpression of SOX9 due to disruption of very-long-range cis-regulatory elements.

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We are grateful to the affected individuals and their families who participated in this study, to the Associations Françaises du Syndrome de Robin, to the Centres de Références Anomalies Cranio-Faciales Rares (AP-HP, Necker and Trousseau hospitals), and to C. Ozilou, G. Staub and G. Guédu-Molina for assistance. We thank T. Attié-Bitach, G. Couly and L. Legeai-Mallet (Necker) and V. van Heyningen and R. Hill (MRC HGU) for useful discussion. This study was underwritten by grants from the Agence Nationale de la Recherche (ERARE grant CraniRare), EUROCRAN FP5, the Fondation pour la Recherche Médicale (FRM), the MRC (UK) and the National Health and Medical Research Council (Australia). S.T. was supported in part by grant NS039818 from the US National Institutes of Health and S.B. by the FRM.

Author information

Author notes

    • Sabina Benko
    • , Judy A Fantes
    • , David R FitzPatrick
    •  & Stanislas Lyonnet

    These authors contributed equally to this work.


  1. INSERM U-781, Hôpital Necker–Enfants Malades, Paris, France.

    • Sabina Benko
    • , Jeanne Amiel
    • , Sophie Thomas
    • , Christelle Golzio
    • , Michel Vekemans
    • , Arnold Munnich
    • , Heather C Etchevers
    • , Anna Pelet
    •  & Stanislas Lyonnet
  2. Medical Research Council Human Genetics Unit (MRC HGU), Institute of Genetic and Molecular Medicine, Edinburgh EH4 2XU, UK.

    • Judy A Fantes
    • , Dirk-Jan Kleinjan
    • , Jacqueline Ramsay
    • , Abdelkader Essafi
    • , Simon Heaney
    • , David McBride
    • , Malcolm Fisher
    • , Paul Perry
    • , Nicholas D Hastie
    •  & David R FitzPatrick
  3. Assistance Publique–Hôpitaux de Paris (AP-HP), Département de Génétique, Hôpital Necker–Enfants Malades, Paris, France.

    • Jeanne Amiel
    • , Michel Vekemans
    • , Arnold Munnich
    •  & Stanislas Lyonnet
  4. Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Australia.

    • Negar Jamshidi
    • , Christopher T Gordon
    • , Nicky Kilpatrick
    • , Paul Thomas
    •  & Peter G Farlie
  5. Université Paris Descartes, Faculté de Médecine, Paris, France.

    • Véronique Abadie
    • , Michel Vekemans
    • , Arnold Munnich
    •  & Stanislas Lyonnet
  6. AP-HP, Service de Pédiatrie, Hôpital Necker–Enfants Malades, Paris, France.

    • Véronique Abadie
  7. Fundación Jiménez Díaz, Genética, Ciberer Madrid, Spain.

    • Carmen Ayuso
  8. CHRU de Lille, Hôpital Jeanne de Flandre, Lille, France.

    • Muriel Holder-Espinasse
  9. North Thames Regional Genetics Service, Great Ormond Street Hospital, London, UK.

    • Melissa M Lees
  10. AP-HP, Service de Chirurgie Maxillo-Faciale et Chirurgie Plastique, Hôpital d'Enfants Armand Trousseau, Paris, France.

    • Arnaud Picard
    •  & Marie-Paule Vazquez
  11. Université Pierre et Marie Curie–Paris 6, UFR de Médecine Pierre et Marie Curie, Paris.

    • Arnaud Picard
    •  & Marie-Paule Vazquez
  12. Wessex Clinical Genetics Academic Group, Division of Human Genetics, University of Southampton, Southampton, UK.

    • I Karen Temple
  13. Department of Biology, École Normale Supérieure, CNRS UMR-8541, Paris, France.

    • Hugues Roest Crollius


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S.B., J.A.F. and A.P. performed molecular genetics studies. J.A.F., C.T.G. and N.J. performed chromosomal studies. S.B. and J.R. performed the in vitro enhancer activity experiments. S.B., S.T., C.G., M.V. and H.C.E. performed human expression studies. J.R., S.B. and A.E. performed immunoprecipitation experiments. D.-J.K. performed transgenic assays. J.A.F., S.H., P.P. and D.B. performed the in vivo chromatin compaction studies. M.F. did the OPT image analysis. S.B. and H.R.C. performed the comparative genomic analysis. J.A., V.A., C.A., M.H.-E., N.K., M.M.L., A.P., I.K.T., M.V., P.T., M.-P.V., D.R.F. and S.L. recruited individuals and families affected with PRS. S.L., D.R.F., P.G.F. and J.A. contributed to the concept, strategy, study design and project management. S.B., H.C.E., S.T., P.G.F., D.R.F. and S.L. contributed to the writing of the manuscript. All authors discussed the results.

Corresponding authors

Correspondence to David R FitzPatrick or Stanislas Lyonnet.

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