Establishment of novel method for introduction of single-nucleotide substitution and application of the method to identification of mutation causing hereditary disease

2014 Fiscal Year Final Research Report

• Project/Area Number: 25830138
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Medical genome science
• Research Institution: Hiroshima University
• Principal Investigator: OCHIAI Hiorhi 広島大学, 理学(系)研究科(研究院), 特任講師 (60640753)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: TALEN / 一塩基多型 / 一塩基置換 / ゲノム編集 / 遺伝子ターゲティング / PCS / 遺伝性疾患

Outline of Final Research Achievements

Premature chromatid separation (PCS) syndrome is a rare autosomal recessive disorder characterized by constitutional aneuploidy and a high risk of childhood cancer. One of the causes of the disease is functional insufficiency of BUBR1 protein. We found a unique single nucleotide substitution 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 TALEN-mediated two-step single-base pair editing strategy and biallelically introduced this substitution into cultured human cells. The cell clones showed reduced BUB1B transcripts, and increased PCS frequency, which are the hallmarks of the syndrome. These results suggested that the nucleotide substitution identified was the causal mutation of PCS syndrome.

Free Research Field

分子生物学