用 2′,3′-二脱氧胞苷高效消除哺乳动物细胞中的 mtDNA

DNA Pub Date : 2024-07-04 DOI:10.3390/dna4030013
N. Kozhukhar, Mikhail F. Alexeyev
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引用次数: 0

摘要

在旨在阐明线粒体DNA对各种细胞过程的贡献或核基因组与线粒体基因组之间相互作用的研究中,不含线粒体DNA(mtDNA)的哺乳动物细胞系是不可或缺的。然而,生成这种细胞(也称为 rho-0 或 ρ0细胞)的工具仍然有限,而且这种方法仍然耗时耗力,最终限制了它们的可用性。最常用于诱导哺乳动物细胞中 mtDNA 缺失的溴化乙锭(EtBr)具有细胞抑制和诱变作用,因为它同时影响核基因组和线粒体基因组。因此,人们对生成ρ0细胞系的新工具越来越感兴趣。在这里,我们研究了 2′,3′-二脱氧胞苷(ddC,扎西他滨)单独使用或与 EtBr 联用生成小鼠和人源ρ0 细胞系以及在小鼠/人体细胞杂交中诱导ρ0 状态的实用性。我们报告说,在永生化小鼠成纤维细胞和人类 143B 细胞中,ddC 都优于 EtBr。此外,与 EtBr 不同的是,ddC 在测试的最高浓度(200 μM)下没有细胞抑制作用,因此更适合普遍使用。我们的结论是,ddC 是生成哺乳动物 ρ0 细胞系的一种很有前途的新工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficient Elimination of mtDNA from Mammalian Cells with 2′,3′-Dideoxycytidine
Mammalian cell lines devoid of mitochondrial DNA (mtDNA) are indispensable in studies aimed at elucidating the contribution of mtDNA to various cellular processes or interactions between nuclear and mitochondrial genomes. However, the repertoire of tools for generating such cells (also known as rho-0 or ρ0 cells) remains limited, and approaches remain time- and labor-intensive, ultimately limiting their availability. Ethidium bromide (EtBr), which is most commonly used to induce mtDNA loss in mammalian cells, is cytostatic and mutagenic as it affects both nuclear and mitochondrial genomes. Therefore, there is growing interest in new tools for generating ρ0 cell lines. Here, we examined the utility of 2′,3′-dideoxycytidine (ddC, zalcitabine) alone or in combination with EtBr for generating ρ0 cell lines of mouse and human origin as well as inducing the ρ0 state in mouse/human somatic cell hybrids. We report that ddC is superior to EtBr in both immortalized mouse fibroblasts and human 143B cells. Also, unlike EtBr, ddC exhibits no cytostatic effects at the highest concentration tested (200 μM), making it more suitable for general use. We conclude that ddC is a promising new tool for generating mammalian ρ0 cell lines.
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DNA
DNA
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