Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient

Nitin Sabherwal, Fiona Bangs, Röth Ralph, Weiss Birgit , Jantz Karin , Tiecke Eva , Hinke Georg , Spaich Christiane , Hauffa Berthold , Kamp Hetty , Kapeller Johannes, Cheryll Tickle, Rappold Gudrun

Research output: Contribution to journalArticle

  • 83 Citations

Abstract

Defects in long-range regulatory elements have recently emerged as previously underestimated factors in the genesis of human congenital disorders. Léri-Weill dyschondrosteosis is a dominant skeletal malformation syndrome caused by mutations in the short stature homeobox gene SHOX. We have analysed four families with Léri-Weill dyschondrosteosis with deletions in the pseudoautosomal region but still with an intact SHOX coding region. Using fluorescence in situ hybridization and single nucleotide polymorphism studies, we identified an interval of ∼200 kb that was deleted in all tested affected family members but retained in the unaffected members and in 100 control individuals. Comparative genomic analysis of this interval revealed eight highly conserved non-genic elements between 48 and 215 kb downstream of the SHOX gene. As mice do not have a Shox gene, we analysed the enhancer potential in chicken embryos using a green fluorescent protein reporter construct driven by the β-globin promoter, by in ovo electroporation of the limb bud. We observed cis-regulatory activity in three of the eight non-genic elements in the developing limbs arguing for an extensive control region of this gene. These findings are consistent with the idea that the deleted region in the affected families contains several distinct elements that regulate Shox expression in the developing limb. Furthermore, the deletion of these elements in humans generates a phenotype apparently undistinguishable to those patients identified with mutations in the SHOX coding region and, for the first time, demonstrates the potential of an in vivo assay in chicken to monitor putative enhancer activity in relation to human disease.
LanguageEnglish
Pages210-222
JournalHuman Molecular Genetics
Volume16
Issue number2
DOIs
StatusPublished - 2007

Fingerprint

Chickens
Extremities
Phenotype
Genes
Limb Buds
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Mutation
beta-Globins
Electroporation
Homeobox Genes
Green Fluorescent Proteins
Fluorescence In Situ Hybridization
Single Nucleotide Polymorphism
Embryonic Structures
Leri-Weil syndrome
Pseudoautosomal Regions

Cite this

Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient. / Sabherwal, Nitin; Bangs, Fiona; Ralph, Röth; Birgit , Weiss; Karin , Jantz; Eva , Tiecke; Georg , Hinke; Christiane , Spaich; Berthold , Hauffa; Hetty , Kamp; Johannes, Kapeller; Tickle, Cheryll; Gudrun , Rappold.

In: Human Molecular Genetics, Vol. 16, No. 2, 2007, p. 210-222.

Research output: Contribution to journalArticle

Sabherwal, N, Bangs, F, Ralph, R, Birgit , W, Karin , J, Eva , T, Georg , H, Christiane , S, Berthold , H, Hetty , K, Johannes, K, Tickle, C & Gudrun , R 2007, 'Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient' Human Molecular Genetics, vol. 16, no. 2, pp. 210-222. DOI: 10.1093/hmg/ddl470
Sabherwal, Nitin ; Bangs, Fiona ; Ralph, Röth ; Birgit , Weiss ; Karin , Jantz ; Eva , Tiecke ; Georg , Hinke ; Christiane , Spaich ; Berthold , Hauffa ; Hetty , Kamp ; Johannes, Kapeller ; Tickle, Cheryll ; Gudrun , Rappold. / Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient. In: Human Molecular Genetics. 2007 ; Vol. 16, No. 2. pp. 210-222
@article{3ec6abea2857447894ddf983e05eea1e,
title = "Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient",
abstract = "Defects in long-range regulatory elements have recently emerged as previously underestimated factors in the genesis of human congenital disorders. L{\'e}ri-Weill dyschondrosteosis is a dominant skeletal malformation syndrome caused by mutations in the short stature homeobox gene SHOX. We have analysed four families with L{\'e}ri-Weill dyschondrosteosis with deletions in the pseudoautosomal region but still with an intact SHOX coding region. Using fluorescence in situ hybridization and single nucleotide polymorphism studies, we identified an interval of ∼200 kb that was deleted in all tested affected family members but retained in the unaffected members and in 100 control individuals. Comparative genomic analysis of this interval revealed eight highly conserved non-genic elements between 48 and 215 kb downstream of the SHOX gene. As mice do not have a Shox gene, we analysed the enhancer potential in chicken embryos using a green fluorescent protein reporter construct driven by the β-globin promoter, by in ovo electroporation of the limb bud. We observed cis-regulatory activity in three of the eight non-genic elements in the developing limbs arguing for an extensive control region of this gene. These findings are consistent with the idea that the deleted region in the affected families contains several distinct elements that regulate Shox expression in the developing limb. Furthermore, the deletion of these elements in humans generates a phenotype apparently undistinguishable to those patients identified with mutations in the SHOX coding region and, for the first time, demonstrates the potential of an in vivo assay in chicken to monitor putative enhancer activity in relation to human disease.",
author = "Nitin Sabherwal and Fiona Bangs and R{\"o}th Ralph and Weiss Birgit and Jantz Karin and Tiecke Eva and Hinke Georg and Spaich Christiane and Hauffa Berthold and Kamp Hetty and Kapeller Johannes and Cheryll Tickle and Rappold Gudrun",
year = "2007",
doi = "10.1093/hmg/ddl470",
language = "English",
volume = "16",
pages = "210--222",
journal = "Human Molecular Genetics",
issn = "0964-6906",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - Long-range conserved non-coding SHOX sequences regulate expression in developing chicken limb and are associated with short stature phenotypes in human patient

AU - Sabherwal,Nitin

AU - Bangs,Fiona

AU - Ralph,Röth

AU - Birgit ,Weiss

AU - Karin ,Jantz

AU - Eva ,Tiecke

AU - Georg ,Hinke

AU - Christiane ,Spaich

AU - Berthold ,Hauffa

AU - Hetty ,Kamp

AU - Johannes,Kapeller

AU - Tickle,Cheryll

AU - Gudrun ,Rappold

PY - 2007

Y1 - 2007

N2 - Defects in long-range regulatory elements have recently emerged as previously underestimated factors in the genesis of human congenital disorders. Léri-Weill dyschondrosteosis is a dominant skeletal malformation syndrome caused by mutations in the short stature homeobox gene SHOX. We have analysed four families with Léri-Weill dyschondrosteosis with deletions in the pseudoautosomal region but still with an intact SHOX coding region. Using fluorescence in situ hybridization and single nucleotide polymorphism studies, we identified an interval of ∼200 kb that was deleted in all tested affected family members but retained in the unaffected members and in 100 control individuals. Comparative genomic analysis of this interval revealed eight highly conserved non-genic elements between 48 and 215 kb downstream of the SHOX gene. As mice do not have a Shox gene, we analysed the enhancer potential in chicken embryos using a green fluorescent protein reporter construct driven by the β-globin promoter, by in ovo electroporation of the limb bud. We observed cis-regulatory activity in three of the eight non-genic elements in the developing limbs arguing for an extensive control region of this gene. These findings are consistent with the idea that the deleted region in the affected families contains several distinct elements that regulate Shox expression in the developing limb. Furthermore, the deletion of these elements in humans generates a phenotype apparently undistinguishable to those patients identified with mutations in the SHOX coding region and, for the first time, demonstrates the potential of an in vivo assay in chicken to monitor putative enhancer activity in relation to human disease.

AB - Defects in long-range regulatory elements have recently emerged as previously underestimated factors in the genesis of human congenital disorders. Léri-Weill dyschondrosteosis is a dominant skeletal malformation syndrome caused by mutations in the short stature homeobox gene SHOX. We have analysed four families with Léri-Weill dyschondrosteosis with deletions in the pseudoautosomal region but still with an intact SHOX coding region. Using fluorescence in situ hybridization and single nucleotide polymorphism studies, we identified an interval of ∼200 kb that was deleted in all tested affected family members but retained in the unaffected members and in 100 control individuals. Comparative genomic analysis of this interval revealed eight highly conserved non-genic elements between 48 and 215 kb downstream of the SHOX gene. As mice do not have a Shox gene, we analysed the enhancer potential in chicken embryos using a green fluorescent protein reporter construct driven by the β-globin promoter, by in ovo electroporation of the limb bud. We observed cis-regulatory activity in three of the eight non-genic elements in the developing limbs arguing for an extensive control region of this gene. These findings are consistent with the idea that the deleted region in the affected families contains several distinct elements that regulate Shox expression in the developing limb. Furthermore, the deletion of these elements in humans generates a phenotype apparently undistinguishable to those patients identified with mutations in the SHOX coding region and, for the first time, demonstrates the potential of an in vivo assay in chicken to monitor putative enhancer activity in relation to human disease.

UR - http://dx.doi.org/10.1093/hmg/ddl470

U2 - 10.1093/hmg/ddl470

DO - 10.1093/hmg/ddl470

M3 - Article

VL - 16

SP - 210

EP - 222

JO - Human Molecular Genetics

T2 - Human Molecular Genetics

JF - Human Molecular Genetics

SN - 0964-6906

IS - 2

ER -