Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines
Summary: Conventional proximity ligation assay (PLA) suffers from target specificity issues that curtail their accuracy on interpreting proximal interactions in cell biology. Here, we present a reliable and sensitive approach by including a fluorochrome-labeled mRNA fragment along with biotin-labele...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722002209 |
| _version_ | 1818208846914519040 |
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| author | Jasmine George Sonam Mittal Ishaque P. Kadamberi Sunila Pradeep Pradeep Chaluvally-Raghavan |
| author_facet | Jasmine George Sonam Mittal Ishaque P. Kadamberi Sunila Pradeep Pradeep Chaluvally-Raghavan |
| author_sort | Jasmine George |
| collection | DOAJ |
| description | Summary: Conventional proximity ligation assay (PLA) suffers from target specificity issues that curtail their accuracy on interpreting proximal interactions in cell biology. Here, we present a reliable and sensitive approach by including a fluorochrome-labeled mRNA fragment along with biotin-labeled RNA probe and a target-specific antibody, which were used to generate proximal ligation signals through linear connectors in intact cells. This protocol will be particularly useful for studying the proximal interactions between RNA binding proteins (RBPs) and their target mRNAs in cells.For complete details on the use and execution of this protocol, please refer to George et al. (2021). |
| first_indexed | 2024-12-12T04:51:19Z |
| format | Article |
| id | doaj.art-be2a8c20427d40fc99b41192d0c746af |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-12T04:51:19Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-be2a8c20427d40fc99b41192d0c746af2022-12-22T00:37:28ZengElsevierSTAR Protocols2666-16672022-06-0132101340Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell linesJasmine George0Sonam Mittal1Ishaque P. Kadamberi2Sunila Pradeep3Pradeep Chaluvally-Raghavan4Department of Obstetrics and Gynecology, Milwaukee, WI 53226, USADepartment of Obstetrics and Gynecology, Milwaukee, WI 53226, USADepartment of Obstetrics and Gynecology, Milwaukee, WI 53226, USADepartment of Obstetrics and Gynecology, Milwaukee, WI 53226, USA; Department of Physiology, Milwaukee, WI 53226, USA; Medical College of Wisconsin Cancer Center, Milwaukee, WI 53226, USADepartment of Obstetrics and Gynecology, Milwaukee, WI 53226, USA; Department of Physiology, Milwaukee, WI 53226, USA; Medical College of Wisconsin Cancer Center, Milwaukee, WI 53226, USA; Corresponding authorSummary: Conventional proximity ligation assay (PLA) suffers from target specificity issues that curtail their accuracy on interpreting proximal interactions in cell biology. Here, we present a reliable and sensitive approach by including a fluorochrome-labeled mRNA fragment along with biotin-labeled RNA probe and a target-specific antibody, which were used to generate proximal ligation signals through linear connectors in intact cells. This protocol will be particularly useful for studying the proximal interactions between RNA binding proteins (RBPs) and their target mRNAs in cells.For complete details on the use and execution of this protocol, please refer to George et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166722002209CancerCell BiologyCell-based AssaysMicroscopyMolecular/Chemical Probes |
| spellingShingle | Jasmine George Sonam Mittal Ishaque P. Kadamberi Sunila Pradeep Pradeep Chaluvally-Raghavan Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines STAR Protocols Cancer Cell Biology Cell-based Assays Microscopy Molecular/Chemical Probes |
| title | Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines |
| title_full | Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines |
| title_fullStr | Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines |
| title_full_unstemmed | Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines |
| title_short | Optimized proximity ligation assay (PLA) for detection of RNA-protein complex interactions in cell lines |
| title_sort | optimized proximity ligation assay pla for detection of rna protein complex interactions in cell lines |
| topic | Cancer Cell Biology Cell-based Assays Microscopy Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722002209 |
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