相关实验视频

Updated: Jul 26, 2025

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Published on: May 10, 2022

以彗星为基础的方法进行比较,以评估基排泄修复

Congying Zheng^1,2,3, Sergey Shaposhnikov^2,3, Andrew Collins²

¹Department of Pharmacology and Toxicology, NUTRIM School of Nutrition and Translational Research in Metabolism, Maastricht University, 6200, Maastricht, Netherlands.

Archives of toxicology

|June 22, 2023

概括

两种用于DNA修复评估的彗星测定方法没有直接的相关性. 细胞修复试验和体外修复试验测量不同的DNA修复终点,这表明它们是互补的工具.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物化学生化学

背景情况:

DNA 修复对于基因组的稳定性至关重要.
彗星测试变种,包括细胞和体外修复测试,用于评估DNA修复.
这些测试评估了DNA修复过程的不同方面.

研究的目的:

直接比较细胞修复试验和体外修复试验.
为了确定这两种基于彗星测试的方法是否测量相同的DNA修复终点.
为了研究DNA切割活动和背景DNA损伤之间的关系.

主要方法:

细胞修复试验和体外修复试验在8个细胞系和人类淋巴细胞中进行的直接比较.
在细胞修复试验中,随着时间的推移评估DNA损伤的消除.
在体外修复试验中评估细胞提取物识别和切割受损DNA的能力.

主要成果:

在细胞修复试验和体外修复试验之间没有发现显著的关系 (R2 = 0.084,P = 0.52).
在体外测试中,DNA切割活动与背景DNA损伤相关 (R2 = 0.621,P = 0.012).
暴露于DNA破坏性物质后,切割活动显著增加,这表明可诱导的基切割修复.

结论:

细胞修复试验和体外修复试验不能测量DNA修复的相同终点.
这两种测试应该被认为是对DNA修复的综合性评估的补充.
试管体内修复试验反映了可诱导的DNA修复机制和背景损伤水平.

关键词:

细胞修复检测试验细胞修复试验彗星测量试验方法修复DNA的修复DNA的修复基因毒性基因毒性在体外修复试验试验.

更多相关视频

Assaying DNA Damage in Hippocampal Neurons Using the Comet Assay

Assaying DNA Damage in Hippocampal Neurons Using the Comet Assay

Published on: December 19, 2012

CometChip: A High-throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells

CometChip: A High-throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells

Published on: October 18, 2014

相关概念视频

Base Excision Repair

Base Excision Repair

One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...

Long-patch Base Excision Repair

Long-patch Base Excision Repair

Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:

Nucleotide Excision Repair

Nucleotide Excision Repair

Base-pairing and DNA Repair

Base-pairing and DNA Repair

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...