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Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy

Site-directed A-to-I RNA editing is a technology for re-programming genetic information at the RNA-level. We describe here the first design of genetically encodable guideRNAs that enable the re-addressing of human ADAR2 toward specific sites in user-defined mRNA targets. Up to 65% editing yield has...

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Detalles Bibliográficos
Autores principales: Wettengel, Jacqueline, Reautschnig, Philipp, Geisler, Sven, Kahle, Philipp J., Stafforst, Thorsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389476/
https://www.ncbi.nlm.nih.gov/pubmed/27907896
http://dx.doi.org/10.1093/nar/gkw911
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author Wettengel, Jacqueline
Reautschnig, Philipp
Geisler, Sven
Kahle, Philipp J.
Stafforst, Thorsten
author_facet Wettengel, Jacqueline
Reautschnig, Philipp
Geisler, Sven
Kahle, Philipp J.
Stafforst, Thorsten
author_sort Wettengel, Jacqueline
collection PubMed
description Site-directed A-to-I RNA editing is a technology for re-programming genetic information at the RNA-level. We describe here the first design of genetically encodable guideRNAs that enable the re-addressing of human ADAR2 toward specific sites in user-defined mRNA targets. Up to 65% editing yield has been achieved in cell culture for the recoding of a premature Stop codon (UAG) into tryptophan (UIG). In the targeted gene, editing was very specific. We applied the technology to recode a recessive loss-of-function mutation in PINK1 (W437X) in HeLa cells and showed functional rescue of PINK1/Parkin-mediated mitophagy, which is linked to the etiology of Parkinson's disease. In contrast to other editing strategies, this approach requires no artificial protein. Our novel guideRNAs may allow for the development of a platform technology that requires only the administration or expression of a guideRNA to recode genetic information, with high potential for application in biology and medicine.
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spelling pubmed-53894762017-04-24 Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy Wettengel, Jacqueline Reautschnig, Philipp Geisler, Sven Kahle, Philipp J. Stafforst, Thorsten Nucleic Acids Res RNA Site-directed A-to-I RNA editing is a technology for re-programming genetic information at the RNA-level. We describe here the first design of genetically encodable guideRNAs that enable the re-addressing of human ADAR2 toward specific sites in user-defined mRNA targets. Up to 65% editing yield has been achieved in cell culture for the recoding of a premature Stop codon (UAG) into tryptophan (UIG). In the targeted gene, editing was very specific. We applied the technology to recode a recessive loss-of-function mutation in PINK1 (W437X) in HeLa cells and showed functional rescue of PINK1/Parkin-mediated mitophagy, which is linked to the etiology of Parkinson's disease. In contrast to other editing strategies, this approach requires no artificial protein. Our novel guideRNAs may allow for the development of a platform technology that requires only the administration or expression of a guideRNA to recode genetic information, with high potential for application in biology and medicine. Oxford University Press 2017-03-17 2016-10-07 /pmc/articles/PMC5389476/ /pubmed/27907896 http://dx.doi.org/10.1093/nar/gkw911 Text en © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://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 RNA
Wettengel, Jacqueline
Reautschnig, Philipp
Geisler, Sven
Kahle, Philipp J.
Stafforst, Thorsten
Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title_full Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title_fullStr Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title_full_unstemmed Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title_short Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
title_sort harnessing human adar2 for rna repair – recoding a pink1 mutation rescues mitophagy
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389476/
https://www.ncbi.nlm.nih.gov/pubmed/27907896
http://dx.doi.org/10.1093/nar/gkw911
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