Targeted Formation of 8-Oxoguanine in Telomeres.
Targeted Formation of 8-Oxoguanine in Telomeres.
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DOI：
10.1007/978-1-0716-2063-2_9
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发表时间：
2022-01-01
期刊：
影响因子：
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通讯作者：
Opresko, Patricia L
中科院分区：
文献类型：
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作者：
Barnes, Ryan P;Thosar, Sanjana A;Opresko, Patricia L
哺乳动物端粒是富含鸟嘌呤的序列，其覆盖线性染色体的末端。虽然被认为是对氧化应激敏感的位点，但对端粒氧化损伤后果的研究主要限于在整个基因组和细胞中引起氧化损伤的实验条件。我们开发了一种化学光遗传学工具（FAP-mCER-TRF 1），以特异性诱导端粒处的单线态氧，从而形成常见的氧化损伤8-氧代鸟嘌呤。在这里，我们描述了这个工具，并详细介绍了如何生成在端粒表达FAP-mCER-TRF 1的细胞系并验证8-氧代-鸟嘌呤的形成。
Mammalian telomeres are guanine-rich sequences which cap the ends of linear chromosomes. While recognized as sites sensitive to oxidative stress, studies on the consequences of oxidative damage to telomeres have been primarily limited to experimental conditions which cause oxidative damage throughout the whole genome and cell. We developed a chemoptogenetic tool (FAP-mCER-TRF1) to specifically induce singlet oxygen at telomeres, resulting in the formation of the common oxidative lesion 8-oxo-guanine. Here, we describe this tool and detail how to generate cell lines which express FAP-mCER-TRF1 at telomeres and verify the formation of 8-oxo-guanine.