30 years of the Comet Assay: an overview with some new insights
By means of this ‘Frontiers in Genetics’ research topic, we are celebrating 30 years of the Comet Assay. The first paper on this single-cell gel electrophoresis assay was published in 1984 by O. Ostling and K.J. Johanson (Biochem. Biophys. Res. Commun. Vol.123: 291-298). The comet assay is a versati...
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| বিন্যাস: | Online |
| ভাষা: | ইংরেজি |
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Frontiers Media SA
2021
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| বিষয়গুলি: | |
| অনলাইন ব্যবহার করুন: | 19552 |
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কোনো ট্যাগ নেই, প্রথমজন হিসাবে ট্যাগ করুন!
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| _version_ | 1869531070026743808 |
|---|---|
| author | Andrew Collins Sabine Langie Amaya Azqueta |
| author_browse | Amaya Azqueta Andrew Collins Sabine Langie |
| author_facet | Andrew Collins Sabine Langie Amaya Azqueta |
| author_sort | Andrew Collins |
| collection | Directory of Open Access Books |
| description | By means of this ‘Frontiers in Genetics’ research topic, we are celebrating 30 years of the Comet Assay. The first paper on this single-cell gel electrophoresis assay was published in 1984 by O. Ostling and K.J. Johanson (Biochem. Biophys. Res. Commun. Vol.123: 291-298). The comet assay is a versatile and sensitive method for measuring single - and double-strand breaks in DNA. By including lesion-specific enzymes in the assay, its range and sensitivity are greatly increased, but it is important to bear in mind that their specificity is not absolute. The comet assay (with and without inclusion of lesion-specific enzymes) is widely used as a biomarker assay in human population studies - primarily to measure DNA damage, but increasingly also to assess the capacity of cells for DNA repair. Ostling and Johanson (Biochem. Biophys. Res. Commun., 1984) were also the first to report experiments to measure DNA repair, by simply following the decrease of DNA damage over time after challenging cells with ionising radiation. However, this approach is time-consuming and laborious as it requires an extended period of cell culture and is therefore not ideal for biomonitoring studies, which typically require high-throughput processing of many samples. As an alternative approach, the in vitro comet-based repair assay was developed: a cell extract is incubated with a DNA substrate containing specific lesions, and DNA incisions accumulate. The in vitro comet-based repair assay has been modified and improved over the past decade: it was first devised to measure base excision repair of oxidised purines in lymphocytes (Collins et al., Mutagenesis, 2001), but has since been adapted for other lesions and thus other repair pathways, as well as being applied to tissue samples in addition to cell suspensions. Even after 30 years, the comet assay is still in a growth phase, with many new users each year. Many questions are repeatedly raised, which may seem to have self-evident answers, but clearly, it is necessary to reiterate them for the benefit of the new audience, and sometimes being forced to think again about old topics can shed new light. Different applications of the comet assay are discussed in this special issue, including: genotoxicity testing in different organisms, human biomonitoring, DNA repair studies, environmental biomonitoring and clinical studies. Furthermore, we consider and where possible answer questions, including the ones raised by Raymond Tice at the 8th International Comet Assay Workshop in Perugia (Italy 2009): What is the spectrum of DNA damage detected by the various versions of the comet assay?; What are the limitations associated with each application?; What should be done to standardize the assay for biomonitoring studies?; Can the comet assay be used to monitor changes in global methylation status?; What cell types are suitable for detecting genotoxic substances and their effects in vivo and in vitro?; Can the assay be fully automated?; and more. So this ‘Frontiers in Genetics’ research topic is written for the beginner as well as for the experienced users of the comet assay. |
| format | Online |
| id | doab-20.500.12854ir-39891 |
| institution | Directory of Open Access Books |
| language | eng |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Frontiers Media SA |
| publisherStr | Frontiers Media SA |
| record_format | ojs |
| spelling | doab-20.500.12854ir-398912024-04-05T12:35:16Z 30 years of the Comet Assay: an overview with some new insights Andrew Collins Sabine Langie Amaya Azqueta QH426-470 Q1-390 Ecotoxicology clinical applications Alkaline comet assay DNA Repair Quality control Genotoxicity testing DNA Damage human biomonitoring high throughput nanotoxicology thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSA Life sciences: general issues::PSAK Genetics (non-medical) By means of this ‘Frontiers in Genetics’ research topic, we are celebrating 30 years of the Comet Assay. The first paper on this single-cell gel electrophoresis assay was published in 1984 by O. Ostling and K.J. Johanson (Biochem. Biophys. Res. Commun. Vol.123: 291-298). The comet assay is a versatile and sensitive method for measuring single - and double-strand breaks in DNA. By including lesion-specific enzymes in the assay, its range and sensitivity are greatly increased, but it is important to bear in mind that their specificity is not absolute. The comet assay (with and without inclusion of lesion-specific enzymes) is widely used as a biomarker assay in human population studies - primarily to measure DNA damage, but increasingly also to assess the capacity of cells for DNA repair. Ostling and Johanson (Biochem. Biophys. Res. Commun., 1984) were also the first to report experiments to measure DNA repair, by simply following the decrease of DNA damage over time after challenging cells with ionising radiation. However, this approach is time-consuming and laborious as it requires an extended period of cell culture and is therefore not ideal for biomonitoring studies, which typically require high-throughput processing of many samples. As an alternative approach, the in vitro comet-based repair assay was developed: a cell extract is incubated with a DNA substrate containing specific lesions, and DNA incisions accumulate. The in vitro comet-based repair assay has been modified and improved over the past decade: it was first devised to measure base excision repair of oxidised purines in lymphocytes (Collins et al., Mutagenesis, 2001), but has since been adapted for other lesions and thus other repair pathways, as well as being applied to tissue samples in addition to cell suspensions. Even after 30 years, the comet assay is still in a growth phase, with many new users each year. Many questions are repeatedly raised, which may seem to have self-evident answers, but clearly, it is necessary to reiterate them for the benefit of the new audience, and sometimes being forced to think again about old topics can shed new light. Different applications of the comet assay are discussed in this special issue, including: genotoxicity testing in different organisms, human biomonitoring, DNA repair studies, environmental biomonitoring and clinical studies. Furthermore, we consider and where possible answer questions, including the ones raised by Raymond Tice at the 8th International Comet Assay Workshop in Perugia (Italy 2009): What is the spectrum of DNA damage detected by the various versions of the comet assay?; What are the limitations associated with each application?; What should be done to standardize the assay for biomonitoring studies?; Can the comet assay be used to monitor changes in global methylation status?; What cell types are suitable for detecting genotoxic substances and their effects in vivo and in vitro?; Can the assay be fully automated?; and more. So this ‘Frontiers in Genetics’ research topic is written for the beginner as well as for the experienced users of the comet assay. 2021-02-11T07:33:57Z 2021-02-11T07:33:57Z 2016-08-16 10:34:25 2015 book 19552 16648714 9782889196494 https://directory.doabooks.org/handle/20.500.12854/39891 eng Frontiers Research Topics image/jpeg Attribution 4.0 International http://www.frontiersin.org/books/30_years_of_the_Comet_Assay_an_overview_with_some_new_insights/675#nogo http://journal.frontiersin.org/researchtopic/1978/30-years-of-the-comet-assay-an-overview-with-some-new-insights Frontiers Media SA 10.3389/978-2-88919-649-4 10.3389/978-2-88919-649-4 bf5ce210-e72e-4860-ba9b-c305640ff3ae 9782889196494 174 open access |
| spellingShingle | QH426-470 Q1-390 Ecotoxicology clinical applications Alkaline comet assay DNA Repair Quality control Genotoxicity testing DNA Damage human biomonitoring high throughput nanotoxicology thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSA Life sciences: general issues::PSAK Genetics (non-medical) Andrew Collins Sabine Langie Amaya Azqueta 30 years of the Comet Assay: an overview with some new insights |
| title | 30 years of the Comet Assay: an overview with some new insights |
| title_full | 30 years of the Comet Assay: an overview with some new insights |
| title_fullStr | 30 years of the Comet Assay: an overview with some new insights |
| title_full_unstemmed | 30 years of the Comet Assay: an overview with some new insights |
| title_short | 30 years of the Comet Assay: an overview with some new insights |
| title_sort | 30 years of the comet assay an overview with some new insights |
| topic | QH426-470 Q1-390 Ecotoxicology clinical applications Alkaline comet assay DNA Repair Quality control Genotoxicity testing DNA Damage human biomonitoring high throughput nanotoxicology thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSA Life sciences: general issues::PSAK Genetics (non-medical) |
| topic_facet | QH426-470 Q1-390 Ecotoxicology clinical applications Alkaline comet assay DNA Repair Quality control Genotoxicity testing DNA Damage human biomonitoring high throughput nanotoxicology thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSA Life sciences: general issues::PSAK Genetics (non-medical) |
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