B P Plitta-Michalak, A Ramos, D Stepien, M Trusiak, M Michalak
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引用次数: 0
摘要
保持核基因组的完整性对于细胞、组织和生物体的活力和整体健康至关重要。DNA 在生理条件下容易受到损伤,并且易受内源性和环境因素的影响,因此面临着持续不断的威胁。为了评估真核细胞内的 DNA 损伤和修复情况,彗星试验是一种基于凝胶电泳、相对简单且灵敏度高的多功能方法。彗星测定法最初设计用于监测哺乳动物细胞群内的 DNA 损伤和修复,现在已在酵母、原生动物、植物和无脊椎动物等多个领域得到应用。这项技术在低温保存研究中被证明是非常有价值的,是确定合适的低温保存方案的重要辅助手段。这些方案包括与低温保护剂、样品制备以及时间和温度方面的储存条件有关的选择。在动物低温保存研究领域,彗星试验是评估 DNA 完整性的黄金标准方法。然而,由于植物细胞的独特性质和相关的技术挑战,在应用于面向植物的研究时,必须做出更多努力。本综述阐明了彗星试验的基本原理,讨论了其当前的迭代,并划分了其在动物和植物标本低温保存中的应用。此外,我们还深入探讨了彗星测定作为植物样本低温保存监测工具所面临的主要挑战。https://doi.org/10.54680/fr24110110112。
The comet assay as a method for assessing dna damage in cryopreserved samples.
The preservation of the nuclear genome's integrity is paramount for the viability and overall health of cells, tissues, and organisms. DNA, being susceptible to damage under physiological conditions and vulnerable to both endogenous and environmental factors, faces constant threats. To assess DNA damage and repair within individual eukaryotic cells, the comet assay presents itself as a versatile, gel electrophoresis-based, relatively simple, and highly sensitive method. Originally designed to monitor DNA damage and repair within populations of mammalian cells, the comet assay has now found applications across diverse domains, including yeast, protozoa, plants, and invertebrates. This technique has proven invaluable in cryopreservation studies, serving as a valuable adjunct for determining suitable cryopreservation protocols. These protocols encompass choices related to cryoprotectants, sample preparation, as well as storage conditions in terms of time and temperature. In the realm of animal cryopreservation research, the comet assay stands as a gold-standard method for assessing DNA integrity. Nevertheless, when applied in plant-oriented investigations, additional efforts are essential due to the distinct nature of plant cells and associated technical challenges. This review elucidates the fundamental principles underlying the comet assay, discusses its current iterations, and delineates its applications in the cryopreservation of both animal and plant specimens. Moreover, we delve into the primary challenges confronting the comet assay's utility as a monitoring tool in the context of plant sample cryopreservation. https://doi.org/10.54680/fr24110110112.
期刊介绍:
A bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.