Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells

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 mi...

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Autores principales:	Ekaterina Royba, Tatsuo Miyamoto, Silvia Natsuko Akutsu, Kosuke Hosoba, Hiroshi Tauchi, Yoshiki Kudo, Satoshi Tashiro, Takashi Yamamoto, Shinya Matsuura
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3661078487974b74b3c4edb7755faa26
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spelling	oai:doaj.org-article:3661078487974b74b3c4edb7755faa262021-12-02T16:06:58ZEvaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells10.1038/s41598-017-06393-82045-2322https://doaj.org/article/3661078487974b74b3c4edb7755faa262017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06393-8https://doaj.org/toc/2045-2322Abstract 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.Ekaterina RoybaTatsuo MiyamotoSilvia Natsuko AkutsuKosuke HosobaHiroshi TauchiYoshiki KudoSatoshi TashiroTakashi YamamotoShinya MatsuuraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ekaterina Royba Tatsuo Miyamoto Silvia Natsuko Akutsu Kosuke Hosoba Hiroshi Tauchi Yoshiki Kudo Satoshi Tashiro Takashi Yamamoto Shinya Matsuura Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
description	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.
format	article
author	Ekaterina Royba Tatsuo Miyamoto Silvia Natsuko Akutsu Kosuke Hosoba Hiroshi Tauchi Yoshiki Kudo Satoshi Tashiro Takashi Yamamoto Shinya Matsuura
author_facet	Ekaterina Royba Tatsuo Miyamoto Silvia Natsuko Akutsu Kosuke Hosoba Hiroshi Tauchi Yoshiki Kudo Satoshi Tashiro Takashi Yamamoto Shinya Matsuura
author_sort	Ekaterina Royba
title	Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
title_short	Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
title_full	Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
title_fullStr	Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
title_full_unstemmed	Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
title_sort	evaluation of atm heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/3661078487974b74b3c4edb7755faa26
work_keys_str_mv	AT ekaterinaroyba evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT tatsuomiyamoto evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT silvianatsukoakutsu evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT kosukehosoba evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT hiroshitauchi evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT yoshikikudo evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT satoshitashiro evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT takashiyamamoto evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells AT shinyamatsuura evaluationofatmheterozygousmutationsunderlyingindividualdifferencesinradiosensitivityusinggenomeeditinginhumanculturedcells
_version_	1718384769058734080

Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells

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