title: Analysis of individual differences in radiosensitivity using genome editing.
abstract: Current standards for radiological protection of the public have been uniformly established. However, individual differences in radiosensitivity are suggested to exist in human populations, which could be caused by nucleotide variants of DNA repair genes. In order to verify if such genetic variants are responsible for individual differences in radiosensitivity, they could be introduced into cultured human cells for evaluation. This strategy would make it possible to analyse the effect of candidate nucleotide variants on individual radiosensitivity, independent of the diverse genetic background. However, efficient gene targeting in cultured human cells is difficult due to the low frequency of homologous recombination (HR) repair. The development of artificial nucleases has enabled efficient HR-mediated genome editing to be performed in cultured human cells. A novel genome editing strategy, 'transcription activator-like effector nuclease (TALEN)-mediated two-step single base pair editing', has been developed, and this was used to introduce a nucleotide variant associated with a chromosomal instability syndrome bi-allelically into cultured human cells to demonstrate that it is the causative mutation. It is proposed that this editing technique will be useful to investigate individual radiosensitivity.
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan shinya@hiroshima-u.ac.jp.
authors.forename: S
authors.initials: S
authors.lastname: Matsuura
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Royba
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.
authors.forename: S N
authors.initials: SN
authors.lastname: Akutsu
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yanagihara
authors.affiliation: Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ochiai
authors.affiliation: Department of Obstetrics and Gynaecology, Graduate School of Biomedical Sciences, Hiroshima University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kudo
authors.affiliation: Department of Cellular Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Tashiro
authors.affiliation: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Miyamoto
mesh_terms: D000072669:Gene Editing / Q000379:methods*; D006801:Humans; D011836:Radiation Tolerance; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics*
chemical_list: D000069896:Transcription Activator-Like Effector Nucleases
keywords: Genome editing; Micronucleus assay; Radiosensitivity; Single nucleotide polymorphism