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...

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Autores principales: Stroppel, Anna S, Latifi, Ngadhnjim, Hanswillemenke, Alfred, Tasakis, Rafail Nikolaos, Papavasiliou, F Nina, Stafforst, Thorsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
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.
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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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