利用2'，3'-双脱氧胞苷通过线粒体DNA耗竭与再增殖对培养细胞中线粒体DNA维持功能的评估
Functional Assessment of Mitochondrial DNA Maintenance by Depletion and Repopulation Using 2',3'-Dideoxycytidine in Cultured Cells.
作者信息
Zsurka Gábor, Trombly Genevieve, Schöler Susanne, Blei Daniel, Kunz Wolfram S
机构信息
Division of Neurochemistry, Institute of Experimental Epileptology and Cognition Research, University of Bonn, Bonn, Germany.
Department of Epileptology, University of Bonn, Bonn, Germany.
出版信息
Methods Mol Biol. 2023;2615:229-240. doi: 10.1007/978-1-0716-2922-2_17.
The manipulation of mitochondrial DNA (mtDNA) copy number in cultured cells, using substances that interfere with DNA replication, is a useful tool to investigate various aspects of mtDNA maintenance. Here we describe the use of 2',3'-dideoxycytidine (ddC) to induce a reversible reduction of mtDNA copy number in human primary fibroblasts and human embryonic kidney (HEK293) cells. Once the application of ddC is stopped, cells depleted for mtDNA attempt to recover normal mtDNA copy numbers. The dynamics of repopulation of mtDNA provide a valuable measure for the enzymatic activity of the mtDNA replication machinery.
利用干扰DNA复制的物质来操纵培养细胞中线粒体DNA（mtDNA）的拷贝数，是研究mtDNA维持各个方面的有用工具。在此，我们描述了使用2'，3'-双脱氧胞苷（ddC）在人原代成纤维细胞和人胚肾（HEK293）细胞中诱导mtDNA拷贝数的可逆减少。一旦停止使用ddC，mtDNA耗尽的细胞会试图恢复正常的mtDNA拷贝数。mtDNA重新填充的动力学为mtDNA复制机制的酶活性提供了有价值的衡量标准。
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