A digital PCR-based protocol to detect and quantify RNA editing events at hotspots
Summary: APOBEC3A, CRISPR programmable RNA base editors, or other enzymes can edit RNA transcripts at specific locations or hotspots. Precise quantification of these RNA-editing events is crucial to determine the activity and efficiency of these enzymes in cells. We have developed a quick method to...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722000284 |
| _version_ | 1818321062490800128 |
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| author | Sunwoo Oh Rémi Buisson |
| author_facet | Sunwoo Oh Rémi Buisson |
| author_sort | Sunwoo Oh |
| collection | DOAJ |
| description | Summary: APOBEC3A, CRISPR programmable RNA base editors, or other enzymes can edit RNA transcripts at specific locations or hotspots. Precise quantification of these RNA-editing events is crucial to determine the activity and efficiency of these enzymes in cells. We have developed a quick method to quantify RNA-editing activity using digital PCR, a sensitive and quantitative technique to detect rare mutations by micro-partitioning bulk PCR reactions. This assay allows rapid absolute quantification of RNA editing events in cell lines or patient samples.For complete details on the use and execution of this protocol, please refer to Jalili et al. (2020) and Oh et al. (2021). |
| first_indexed | 2024-12-13T10:34:56Z |
| format | Article |
| id | doaj.art-859939fd04ea434fa892e021341aceeb |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-13T10:34:56Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-859939fd04ea434fa892e021341aceeb2022-12-21T23:50:45ZengElsevierSTAR Protocols2666-16672022-03-0131101148A digital PCR-based protocol to detect and quantify RNA editing events at hotspotsSunwoo Oh0Rémi Buisson1Department of Biological Chemistry, School of Medicine, University of California Irvine, Irvine, CA 92697, USA; Chao Family Comprehensive Cancer Center, University of California Irvine, Irvine, CA 92697, USADepartment of Biological Chemistry, School of Medicine, University of California Irvine, Irvine, CA 92697, USA; Chao Family Comprehensive Cancer Center, University of California Irvine, Irvine, CA 92697, USA; Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University of California Irvine, Irvine, CA 92697, USA; Corresponding authorSummary: APOBEC3A, CRISPR programmable RNA base editors, or other enzymes can edit RNA transcripts at specific locations or hotspots. Precise quantification of these RNA-editing events is crucial to determine the activity and efficiency of these enzymes in cells. We have developed a quick method to quantify RNA-editing activity using digital PCR, a sensitive and quantitative technique to detect rare mutations by micro-partitioning bulk PCR reactions. This assay allows rapid absolute quantification of RNA editing events in cell lines or patient samples.For complete details on the use and execution of this protocol, please refer to Jalili et al. (2020) and Oh et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166722000284CancerCell BiologyCRISPRHealth SciencesMolecular Biology |
| spellingShingle | Sunwoo Oh Rémi Buisson A digital PCR-based protocol to detect and quantify RNA editing events at hotspots STAR Protocols Cancer Cell Biology CRISPR Health Sciences Molecular Biology |
| title | A digital PCR-based protocol to detect and quantify RNA editing events at hotspots |
| title_full | A digital PCR-based protocol to detect and quantify RNA editing events at hotspots |
| title_fullStr | A digital PCR-based protocol to detect and quantify RNA editing events at hotspots |
| title_full_unstemmed | A digital PCR-based protocol to detect and quantify RNA editing events at hotspots |
| title_short | A digital PCR-based protocol to detect and quantify RNA editing events at hotspots |
| title_sort | digital pcr based protocol to detect and quantify rna editing events at hotspots |
| topic | Cancer Cell Biology CRISPR Health Sciences Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722000284 |
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