Analysis of individual differences in radiosensitivity using genome editing
Analysis of individual differences in radiosensitivity using genome editing
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DOI：
10.1177/0146645316633941
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发表时间：
2016-06-01
影响因子：
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通讯作者：
Miyamoto, T.
中科院分区：
文献类型：
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作者：
Matsuura, S.;Royba, E.;Miyamoto, T.
统一制定了现行的公众辐射防护标准。然而，辐射敏感性的个体差异被认为存在于人群中，这可能是由DNA修复基因的核苷酸变异引起的。为了验证这些基因变异是否导致了放射敏感性的个体差异，可以将它们引入培养的人类细胞中进行评估。这一策略将使分析候选核苷酸变异对个体放射敏感性的影响成为可能，而不依赖于不同的遗传背景。然而，由于同源重组（HR）修复的频率较低，在培养的人类细胞中进行有效的基因靶向是困难的。人工核酸酶的发展使得在培养的人类细胞中进行有效的hr介导的基因组编辑成为可能。一种新的基因组编辑策略，转录激活因子样效应核酸酶（TALEN）介导的两步单碱基对编辑，已经被开发出来，并被用于将与染色体不稳定性综合征相关的核苷酸变异双等位地引入培养的人类细胞中，以证明它是致病突变。有人提出，这种编辑技术将有助于研究个体的放射敏感性。
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.