title: TALEN-mediated single-base-pair editing identification of an intergenic mutation upstream of BUB1B as causative of PCS (MVA) syndrome.
abstract: Cancer-prone syndrome of premature chromatid separation with mosaic variegated aneuploidy [PCS (MVA) syndrome] is a rare autosomal recessive disorder characterized by constitutional aneuploidy and a high risk of childhood cancer. We previously reported monoallelic mutations in the BUB1B gene (encoding BUBR1) in seven Japanese families with the syndrome. No second mutation was found in the opposite allele of any of the families studied, although a conserved BUB1B haplotype and a decreased transcript were identified. To clarify the molecular pathology of the second allele, we extended our mutational search to a candidate region surrounding BUB1B. A unique single nucleotide substitution, G > A at ss802470619, was identified in an intergenic region 44 kb upstream of a BUB1B transcription start site, which cosegregated with the disorder. To examine whether this is the causal mutation, we designed a transcription activator-like effector nuclease-mediated two-step single-base pair editing strategy and biallelically introduced this substitution into cultured human cells. The cell clones showed reduced BUB1B transcripts, increased PCS frequency, and MVA, which are the hallmarks of the syndrome. We also encountered a case of a Japanese infant with PCS (MVA) syndrome carrying a homozygous single nucleotide substitution at ss802470619. These results suggested that the nucleotide substitution identified was the causal mutation of PCS (MVA) syndrome.
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ochiai
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Miyamoto
authors.forename: Akinori
authors.initials: A
authors.lastname: Kanai
authors.forename: Kosuke
authors.initials: K
authors.lastname: Hosoba
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Sakuma
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Kudo
authors.forename: Keiko
authors.initials: K
authors.lastname: Asami
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ogawa
authors.forename: Akihiro
authors.initials: A
authors.lastname: Watanabe
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kajii
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Shinya
authors.initials: S
authors.lastname: Matsuura
mesh_terms: D000818:Animals; D020029:Base Pairing; D018797:Cell Cycle Proteins; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D013577:Syndrome
chemical_list: C584279:BUB1B protein, human; D018797:Cell Cycle Proteins; D017346:Protein Serine-Threonine Kinases
keywords: genome editing; spindle assembly checkpoint