RRC ID |
50506
|
著者 |
Royba E, Miyamoto T, Natsuko Akutsu S, Hosoba K, Tauchi H, Kudo Y, Tashiro S, Yamamoto T, Matsuura S.
|
タイトル |
Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells.
|
ジャーナル |
Sci Rep
|
Abstract |
Ionizing radiation (IR) induces DNA double-strand breaks (DSBs), which are an initial step towards chromosomal aberrations and cell death. It has been suggested that there are individual differences in radiosensitivity within human populations, and that the variations in DNA repair genes might determine this heterogeneity. However, it is difficult to quantify the effect of genetic variants on the individual differences in radiosensitivity, since confounding factors such as smoking and the diverse genetic backgrounds within human populations affect radiosensitivity. To precisely quantify the effect of a genetic variation on radiosensitivity, we here used the CRISPR-ObLiGaRe (Obligate Ligation-Gated Recombination) method combined with the CRISPR/Cas9 system and a nonhomologous end joining (NHEJ)-mediated knock-in technique in human cultured cells with a uniform genetic background. We generated ATM heterozygous knock-out (ATM +/-) cell clones as a carrier model of a radiation-hypersensitive autosomal-recessive disorder, ataxia-telangiectasia (A-T). Cytokinesis-blocked micronucleus assay and chromosome aberration assay showed that the radiosensitivity of ATM +/- cell clones was significantly higher than that of ATM +/+ cells, suggesting that ATM gene variants are indeed involved in determining individual radiosensitivity. Importantly, the differences in radiosensitivity among the same genotype clones were small, unlike the individual differences in fibroblasts derived from A-T-affected family members.
|
巻・号 |
7(1)
|
ページ |
5996
|
公開日 |
2017-7-20
|
DOI |
10.1038/s41598-017-06393-8
|
PII |
10.1038/s41598-017-06393-8
|
PMID |
28729543
|
PMC |
PMC5519549
|
MeSH |
Ataxia Telangiectasia Mutated Proteins / genetics*
Automation
CRISPR-Cas Systems / genetics
Cells, Cultured
Clone Cells
Cytokinesis
Fibroblasts / metabolism
Fibroblasts / pathology
Gene Editing*
Heterozygote
Humans
Individuality*
Micronucleus Tests
Models, Biological
Mutation / genetics*
Radiation Tolerance / genetics*
Recombination, Genetic / genetics
|
IF |
3.998
|
引用数 |
12
|
リソース情報 |
遺伝子材料 |
pX330-U6-Chimeric_BB-CBh-hSpCas9 sgRNA ATM exon11 (RDB15816) |