Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing
The SNAP-ADAR tool enables precise and efficient A-to-I RNA editing in a guideRNA-dependent manner by applying the self-labeling SNAP-tag enzyme to generate RNA-guided editases in cell culture. Here, we extend this platform by combining the SNAP-tagged tool with further effectors steered by the orth...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450088/ https://www.ncbi.nlm.nih.gov/pubmed/34197596 http://dx.doi.org/10.1093/nar/gkab541 |
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author | Stroppel, Anna S Latifi, Ngadhnjim Hanswillemenke, Alfred Tasakis, Rafail Nikolaos Papavasiliou, F Nina Stafforst, Thorsten |
author_facet | Stroppel, Anna S Latifi, Ngadhnjim Hanswillemenke, Alfred Tasakis, Rafail Nikolaos Papavasiliou, F Nina Stafforst, Thorsten |
author_sort | Stroppel, Anna S |
collection | PubMed |
description | The SNAP-ADAR tool enables precise and efficient A-to-I RNA editing in a guideRNA-dependent manner by applying the self-labeling SNAP-tag enzyme to generate RNA-guided editases in cell culture. Here, we extend this platform by combining the SNAP-tagged tool with further effectors steered by the orthogonal HALO-tag. Due to their small size (ca. 2 kb), both effectors are readily integrated into one genomic locus. We demonstrate selective and concurrent recruitment of ADAR1 and ADAR2 deaminase activity for optimal editing with extended substrate scope and moderate global off-target effects. Furthermore, we combine the recruitment of ADAR1 and APOBEC1 deaminase activity to achieve selective and concurrent A-to-I and C-to-U RNA base editing of endogenous transcripts inside living cells, again with moderate global off-target effects. The platform should be readily transferable to further epitranscriptomic writers and erasers to manipulate epitranscriptomic marks in a programmable way with high molecular precision. |
format | Online Article Text |
id | pubmed-8450088 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-84500882021-09-20 Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing Stroppel, Anna S Latifi, Ngadhnjim Hanswillemenke, Alfred Tasakis, Rafail Nikolaos Papavasiliou, F Nina Stafforst, Thorsten Nucleic Acids Res Methods Online The SNAP-ADAR tool enables precise and efficient A-to-I RNA editing in a guideRNA-dependent manner by applying the self-labeling SNAP-tag enzyme to generate RNA-guided editases in cell culture. Here, we extend this platform by combining the SNAP-tagged tool with further effectors steered by the orthogonal HALO-tag. Due to their small size (ca. 2 kb), both effectors are readily integrated into one genomic locus. We demonstrate selective and concurrent recruitment of ADAR1 and ADAR2 deaminase activity for optimal editing with extended substrate scope and moderate global off-target effects. Furthermore, we combine the recruitment of ADAR1 and APOBEC1 deaminase activity to achieve selective and concurrent A-to-I and C-to-U RNA base editing of endogenous transcripts inside living cells, again with moderate global off-target effects. The platform should be readily transferable to further epitranscriptomic writers and erasers to manipulate epitranscriptomic marks in a programmable way with high molecular precision. Oxford University Press 2021-07-01 /pmc/articles/PMC8450088/ /pubmed/34197596 http://dx.doi.org/10.1093/nar/gkab541 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Methods Online Stroppel, Anna S Latifi, Ngadhnjim Hanswillemenke, Alfred Tasakis, Rafail Nikolaos Papavasiliou, F Nina Stafforst, Thorsten Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title | Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title_full | Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title_fullStr | Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title_full_unstemmed | Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title_short | Harnessing self-labeling enzymes for selective and concurrent A-to-I and C-to-U RNA base editing |
title_sort | harnessing self-labeling enzymes for selective and concurrent a-to-i and c-to-u rna base editing |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450088/ https://www.ncbi.nlm.nih.gov/pubmed/34197596 http://dx.doi.org/10.1093/nar/gkab541 |
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