Gureev Artem P, Nesterova Veronika V, Sadovnikova Irina S
Departments of Genetics, Cytology and Bioengineering, Voronezh State University, Voronezh, Russia.
Departments of Genetics, Cytology and Bioengineering, Voronezh State University, Voronezh, Russia.
DNA Repair (Amst). 2025 Feb;146:103812. doi: 10.1016/j.dnarep.2025.103812. Epub 2025 Jan 17.
Mitochondrial DNA (mtDNA) is often more susceptible to damage compared to nuclear DNA. This is due to its localization in the mitochondrial matrix, where a large portion of reactive oxygen species are produced. Mitochondria do not have histones and mtDNA is only slightly protected by histone-like proteins and is believed to have less efficient repair mechanisms. In this review, we discuss the long-range PCR method, which allows for the effective detection of mtDNA damage. The method is based on the assumption that various types of DNA lesions can interfere the progress of DNA polymerase, resulting in reduced amplification efficiency. It can be used to estimate the number of additional (above background) lesions in mtDNA. The review outlines the evolution of the methodology, its variations, applications in a wide range of model organisms, the advantages of the method and its limitations, as well as ways to overcome these limitations. Over the past two decades, the use of long-range PCR has allowed the study of mtDNA repair mechanisms, the characteristics of mitochondrial genome damage in various neurodegenerative diseases, aging, ischemic and oncological processes, as well as in anticancer therapy. The assessment of mtDNA damage has also been proposed for use in environmental biomonitoring. This review provides a critical evaluation of the various variations of this method, summarizes the accumulated data, and discusses the role of mtDNA damage in different organs at the organismal level.
与核DNA相比，线粒体DNA（mtDNA）通常更容易受到损伤。这是由于其定位于线粒体基质中，而线粒体基质是大量活性氧产生的地方。线粒体没有组蛋白，mtDNA仅受到类组蛋白的轻微保护，并且据信其修复机制效率较低。在本综述中，我们讨论了长程PCR方法，该方法能够有效检测mtDNA损伤。该方法基于这样的假设，即各种类型的DNA损伤会干扰DNA聚合酶的进程，导致扩增效率降低。它可用于估计mtDNA中额外（高于背景）损伤的数量。本综述概述了该方法的演变、其变体、在广泛的模式生物中的应用、该方法的优点及其局限性，以及克服这些局限性的方法。在过去二十年中，长程PCR的使用使得对mtDNA修复机制、各种神经退行性疾病、衰老、缺血和肿瘤过程以及抗癌治疗中线粒体基因组损伤的特征进行了研究。mtDNA损伤评估也已被提议用于环境生物监测。本综述对该方法的各种变体进行了批判性评估，总结了积累的数据，并在机体水平上讨论了mtDNA损伤在不同器官中的作用。
