Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells
| Autor: | Natalie R. Gassman, Joel Andrews, Nathaniel W Holton |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cancer Research Materials science XRCC1 DNA Repair DNA damage DNA repair Confocal General Chemical Engineering CHO Cells General Biochemistry Genetics and Molecular Biology law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cricetulus law Animals Humans DNA Breaks Double-Stranded Irradiation DNA Breaks Single-Stranded laser micro-irradiation Genetics General Immunology and Microbiology Lasers General Neuroscience strand break strand break signaling DNA Laser Double Strand Break Repair gamma-H2AX 030104 developmental biology chemistry 030220 oncology & carcinogenesis Issue 127 Biophysics |
| Zdroj: | Journal of Visualized Experiments : JoVE |
| ISSN: | 1940-087X |
| Popis: | Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular biology techniques have clarified structure, enzymatic functions, and kinetics of repair proteins, there is still a need to understand how repair is coordinated within the nucleus. Laser micro-irradiation offers a powerful tool for inducing DNA damage and monitoring the recruitment of repair proteins. Induction of DNA damage by laser micro-irradiation can occur with a range of wavelengths, and users can reliably induce single strand breaks, base lesions and double strand breaks with a range of doses. Here, laser micro-irradiation is used to examine repair of single and double strand breaks induced by two common confocal laser wavelengths, 355 nm and 405 nm. Further, proper characterization of the applied laser dose for inducing specific damage mixtures is described, so users can reproducibly perform laser micro-irradiation data acquisition and analysis. |
| Databáze: | OpenAIRE |
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