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Updated: Apr 11, 2026
CometChip: A High-throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells
Published on: October 18, 2014
Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and
Xue Zhang1, Chong Zhang, Qian-Qian Zhou
1Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical Engineering, Tsinghua University, Beijing, China.
Abstract:
DNA damage is the dominant source of mutation, which is the driving force of evolution. Therefore, it is important to quantitatively analyze the DNA damage caused by different mutagenesis methods, the subsequent mutation rates, and their relationship. Atmospheric and room temperature plasma (ARTP) mutagenesis has been used for the mutation breeding of more than 40 microorganisms. However, ARTP mutagenesis has not been quantitatively compared with conventional mutation methods. In this study, the umu test using a flow-cytometric analysis was developed to quantify the DNA damage in individual viable cells using Salmonella typhimurium NM2009 as the model strain and to determine the mutation rate. The newly developed method was used to evaluate four different mutagenesis systems: a new ARTP tool, ultraviolet radiation, 4-nitroquinoline-1-oxide (4-NQO), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) mutagenesis. The mutation rate was proportional to the corresponding SOS response induced by DNA damage. ARTP caused greater DNA damage to individual living cells than the other conventional mutagenesis methods, and the mutation rate was also higher. By quantitatively comparing the DNA damage and consequent mutation rate after different types of mutagenesis, we have shown that ARTP is a potentially powerful mutagenesis tool with which to improve the characteristics of microbial cell factories.
Insights
Atmospheric and room temperature plasma (ARTP) mutagenesis causes significant DNA damage and higher mutation rates compared to conventional methods. This study quantifies DNA damage and mutation rates, highlighting ARTP as a potent tool for microbial strain improvement.
Area of Science:
- Microbiology
- Molecular Biology
- Evolutionary Biology
Background:
- DNA damage is a primary driver of evolutionary mutation.
- Quantitative analysis of DNA damage and mutation rates from various mutagenesis methods is crucial.
- Atmospheric and room temperature plasma (ARTP) mutagenesis is used in microbial breeding but lacks quantitative comparison.
Purpose of the Study:
- To develop a quantitative method for assessing DNA damage and mutation rates in individual cells.
- To compare the efficacy of ARTP mutagenesis against conventional methods (UV, 4-NQO, MNNG).
- To evaluate ARTP as a tool for microbial cell factory improvement.
Main Methods:
- Development of a umu test with flow-cytometric analysis for quantifying DNA damage in viable cells.
- Utilizing Salmonella typhimurium NM2009 as a model strain.
- Evaluation of four mutagenesis systems: ARTP, UV, 4-NQO, and MNNG.
Main Results:
- A direct correlation was observed between DNA damage (SOS response) and mutation rate across systems.
- ARTP induced greater DNA damage in individual cells compared to UV, 4-NQO, and MNNG.
- ARTP resulted in a higher mutation rate than the conventional methods tested.
Conclusions:
- The developed flow-cytometric umu test provides a quantitative measure of DNA damage and mutation.
- ARTP mutagenesis demonstrates superior DNA-damaging capacity and mutation induction compared to conventional methods.
- ARTP is a promising mutagenesis tool for enhancing microbial characteristics in industrial applications.
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