Detecting RNA-protein proximity at DNA double-strand breaks using combined fluorescence in situ hybridization with proximity ligation assay
<p>RNA transcribed at DNA double-strand breaks (DSBs) contributes to accurate DNA repair. Here, using the repair factors 53BP1 and TIRR as examples, we combine the fluorescence <i>in situ</i> hybridization (FISH) and proximity ligation assay (PLA) techniques to determine protein pr...
| Main Authors: | , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Cell Press
2023
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| Summary: | <p>RNA transcribed at DNA double-strand breaks (DSBs) contributes to accurate DNA repair. Here, using the repair factors 53BP1 and TIRR as examples, we combine the fluorescence <i>in situ</i> hybridization (FISH) and proximity ligation assay (PLA) techniques to determine protein proximity to DSB-transcribed RNA. In this FISH-PLA protocol, we detail steps for designing DNA probes and image analysis using CellProfiler™ software. This approach has many potential applications for the study of the RNA-binding proteins and nascent RNA interactions.<br>
For complete details on the use and execution of this protocol, please refer to Ketley et al. (2022).<sup>1</sup></p> |
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