title: Deep sequencing of a candidate region harboring the SOX9 gene for the canine XX disorder of sex development.
abstract: A disorder of sex development (DSD) in dogs with female sex chromosomes (78, XX), a lack of the SRY gene and the presence of testes or ovotestes is commonly diagnosed in numerous breeds. The molecular background of DSD is not fully recognized but has been linked to the copy number variation in the region harboring the SOX9 gene. We applied a genome-wide association study and targeted next-generation sequencing techniques to compare DSD and normal female dogs. The genome-wide association study did not indicate a significant chromosome region. Targeted next-generation sequencing of a 1.5-Mb region on canine chromosome 9 harboring the SOX9 gene revealed two putatively DSD-associated copy number variations 355 kb upstream and 691 kb downstream of SOX9, four blocks of low polymorphism and two blocks of an elevated heterozygosity. An initial next-generation sequencing analysis showed an association with two SNPs, but validation in larger cohorts did not confirm this result. We identified a large homologous fragment (over 243.8 kb), named hfMAGI2, located upstream of SOX9, that overlaps a known copy number variation region. It shows a high sequence similarity with the 5' flanking region of the MAGI2 gene located on canine chromosome 18 that encodes a protein involved in ovary formation during early embryonic development. Our study showed that the identified copy number variation region located upstream of the SOX9 gene contains potential regulatory sequences (long non-coding RNA and hfMAGI2) and led to the assumption that a multiplication of this element may alter expression of the SOX9 gene, triggering the DSD phenotype.
authors.affiliation: Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznan, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Nowacka-Woszuk
authors.affiliation: Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznan, Poland.
authors.forename: I
authors.initials: I
authors.lastname: Szczerbal
authors.affiliation: Chair of Animal Breeding, Technische Universitat Munchen, Liesel-Beckmann-Straße 1, D-85354, Freising-Weihenstephan, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Pausch
authors.affiliation: Department of Veterinary Biosciences, University of Helsinki, Helsinki, 00014, Finland.
authors.forename: S
authors.initials: S
authors.lastname: Hundi
authors.affiliation: Department of Veterinary Biosciences, University of Helsinki, Helsinki, 00014, Finland.
authors.forename: M K
authors.initials: MK
authors.lastname: Hytönen
authors.affiliation: Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznan, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Grzemski
authors.affiliation: Chair of Livestock Biotechnology, Technische Universitat Munchen, Liesel-Beckmann-Straße 1, D-85354, Freising-Weihenstephan, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Flisikowski
authors.affiliation: Department of Veterinary Biosciences, University of Helsinki, Helsinki, 00014, Finland.
authors.forename: H
authors.initials: H
authors.lastname: Lohi
authors.affiliation: Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznan, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Switonski
authors.affiliation: Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznan, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Szydlowski
mesh_terms: D000818:Animals; D056915:DNA Copy Number Variations; D012734:Disorders of Sex Development / Q000235:genetics / Q000662:veterinary*; D004283:Dog Diseases / Q000235:genetics*; D004285:Dogs / Q000235:genetics*; D005260:Female; D055106:Genome-Wide Association Study; D059014:High-Throughput Nucleotide Sequencing; D020641:Polymorphism, Single Nucleotide; D055758:SOX9 Transcription Factor / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: D055758:SOX9 Transcription Factor
keywords: 
  MAGI2
  ; RevSex; copy number variation; disorder of sex development; dog; intersexuality; lincRNA