DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation
Accurately characterizing DNA double-stranded breaks (DSBs) and understanding the DNA damage response (DDR) is crucial for assessing cellular genotoxicity, maintaining genomic integrity, and advancing gene editing technologies. Immunofluorescence-based techniques have proven to be invaluable for qua...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/25/4/2227 |
| _version_ | 1797297983133319168 |
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| author | Jake Atkinson Eva Bezak Hien Le Ivan Kempson |
| author_facet | Jake Atkinson Eva Bezak Hien Le Ivan Kempson |
| author_sort | Jake Atkinson |
| collection | DOAJ |
| description | Accurately characterizing DNA double-stranded breaks (DSBs) and understanding the DNA damage response (DDR) is crucial for assessing cellular genotoxicity, maintaining genomic integrity, and advancing gene editing technologies. Immunofluorescence-based techniques have proven to be invaluable for quantifying and visualizing DSB repair, providing valuable insights into cellular repair processes. However, the selection of appropriate markers for analysis can be challenging due to the intricate nature of DSB repair mechanisms, often leading to ambiguous interpretations. This comprehensively summarizes the significance of immunofluorescence-based techniques, with their capacity for spatiotemporal visualization, in elucidating complex DDR processes. By evaluating the strengths and limitations of different markers, we identify where they are most relevant chronologically from DSB detection to repair, better contextualizing what each assay represents at a molecular level. This is valuable for identifying biases associated with each assay and facilitates accurate data interpretation. This review aims to improve the precision of DSB quantification, deepen the understanding of DDR processes, assay biases, and pathway choices, and provide practical guidance on marker selection. Each assay offers a unique perspective of the underlying processes, underscoring the need to select markers that are best suited to specific research objectives. |
| first_indexed | 2024-03-07T22:28:15Z |
| format | Article |
| id | doaj.art-08196f4838f54696a32e6a4507797293 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-07T22:28:15Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-08196f4838f54696a32e6a45077972932024-02-23T15:20:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01254222710.3390/ijms25042227DNA Double Strand Break and Response Fluorescent Assays: Choices and InterpretationJake Atkinson0Eva Bezak1Hien Le2Ivan Kempson3Future Industries Institute, University of South Australia, Mawson Lakes, SA 5095, AustraliaUniSA Allied Health and Human Performance, University of South Australia, Adelaide, SA 5095, AustraliaUniSA Allied Health and Human Performance, University of South Australia, Adelaide, SA 5095, AustraliaFuture Industries Institute, University of South Australia, Mawson Lakes, SA 5095, AustraliaAccurately characterizing DNA double-stranded breaks (DSBs) and understanding the DNA damage response (DDR) is crucial for assessing cellular genotoxicity, maintaining genomic integrity, and advancing gene editing technologies. Immunofluorescence-based techniques have proven to be invaluable for quantifying and visualizing DSB repair, providing valuable insights into cellular repair processes. However, the selection of appropriate markers for analysis can be challenging due to the intricate nature of DSB repair mechanisms, often leading to ambiguous interpretations. This comprehensively summarizes the significance of immunofluorescence-based techniques, with their capacity for spatiotemporal visualization, in elucidating complex DDR processes. By evaluating the strengths and limitations of different markers, we identify where they are most relevant chronologically from DSB detection to repair, better contextualizing what each assay represents at a molecular level. This is valuable for identifying biases associated with each assay and facilitates accurate data interpretation. This review aims to improve the precision of DSB quantification, deepen the understanding of DDR processes, assay biases, and pathway choices, and provide practical guidance on marker selection. Each assay offers a unique perspective of the underlying processes, underscoring the need to select markers that are best suited to specific research objectives.https://www.mdpi.com/1422-0067/25/4/2227microscopynon-homologous end joining (NHEJ)homologous-recombination (HR)γH2AXp53 Binding Protein 1 (53BP1)foci |
| spellingShingle | Jake Atkinson Eva Bezak Hien Le Ivan Kempson DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation International Journal of Molecular Sciences microscopy non-homologous end joining (NHEJ) homologous-recombination (HR) γH2AX p53 Binding Protein 1 (53BP1) foci |
| title | DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation |
| title_full | DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation |
| title_fullStr | DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation |
| title_full_unstemmed | DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation |
| title_short | DNA Double Strand Break and Response Fluorescent Assays: Choices and Interpretation |
| title_sort | dna double strand break and response fluorescent assays choices and interpretation |
| topic | microscopy non-homologous end joining (NHEJ) homologous-recombination (HR) γH2AX p53 Binding Protein 1 (53BP1) foci |
| url | https://www.mdpi.com/1422-0067/25/4/2227 |
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