Cargando…
Prime editing for functional repair in patient-derived disease models
Prime editing is a recent genome editing technology using fusion proteins of Cas9-nickase and reverse transcriptase, that holds promise to correct the vast majority of genetic defects. Here, we develop prime editing for primary adult stem cells grown in organoid culture models. First, we generate pr...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584657/ https://www.ncbi.nlm.nih.gov/pubmed/33097693 http://dx.doi.org/10.1038/s41467-020-19136-7 |
_version_ | 1783599641569263616 |
---|---|
author | Schene, Imre F. Joore, Indi P. Oka, Rurika Mokry, Michal van Vugt, Anke H. M. van Boxtel, Ruben van der Doef, Hubert P. J. van der Laan, Luc J. W. Verstegen, Monique M. A. van Hasselt, Peter M. Nieuwenhuis, Edward E. S. Fuchs, Sabine A. |
author_facet | Schene, Imre F. Joore, Indi P. Oka, Rurika Mokry, Michal van Vugt, Anke H. M. van Boxtel, Ruben van der Doef, Hubert P. J. van der Laan, Luc J. W. Verstegen, Monique M. A. van Hasselt, Peter M. Nieuwenhuis, Edward E. S. Fuchs, Sabine A. |
author_sort | Schene, Imre F. |
collection | PubMed |
description | Prime editing is a recent genome editing technology using fusion proteins of Cas9-nickase and reverse transcriptase, that holds promise to correct the vast majority of genetic defects. Here, we develop prime editing for primary adult stem cells grown in organoid culture models. First, we generate precise in-frame deletions in the gene encoding β‐catenin (CTNNB1) that result in proliferation independent of Wnt-stimuli, mimicking a mechanism of the development of liver cancer. Moreover, prime editing functionally recovers disease-causing mutations in intestinal organoids from patients with DGAT1-deficiency and liver organoids from a patient with Wilson disease (ATP7B). Prime editing is as efficient in 3D grown organoids as in 2D grown cell lines and offers greater precision than Cas9-mediated homology directed repair (HDR). Base editing remains more reliable than prime editing but is restricted to a subgroup of pathogenic mutations. Whole-genome sequencing of four prime-edited clonal organoid lines reveals absence of genome-wide off-target effects underscoring therapeutic potential of this versatile and precise gene editing strategy. |
format | Online Article Text |
id | pubmed-7584657 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-75846572020-10-29 Prime editing for functional repair in patient-derived disease models Schene, Imre F. Joore, Indi P. Oka, Rurika Mokry, Michal van Vugt, Anke H. M. van Boxtel, Ruben van der Doef, Hubert P. J. van der Laan, Luc J. W. Verstegen, Monique M. A. van Hasselt, Peter M. Nieuwenhuis, Edward E. S. Fuchs, Sabine A. Nat Commun Article Prime editing is a recent genome editing technology using fusion proteins of Cas9-nickase and reverse transcriptase, that holds promise to correct the vast majority of genetic defects. Here, we develop prime editing for primary adult stem cells grown in organoid culture models. First, we generate precise in-frame deletions in the gene encoding β‐catenin (CTNNB1) that result in proliferation independent of Wnt-stimuli, mimicking a mechanism of the development of liver cancer. Moreover, prime editing functionally recovers disease-causing mutations in intestinal organoids from patients with DGAT1-deficiency and liver organoids from a patient with Wilson disease (ATP7B). Prime editing is as efficient in 3D grown organoids as in 2D grown cell lines and offers greater precision than Cas9-mediated homology directed repair (HDR). Base editing remains more reliable than prime editing but is restricted to a subgroup of pathogenic mutations. Whole-genome sequencing of four prime-edited clonal organoid lines reveals absence of genome-wide off-target effects underscoring therapeutic potential of this versatile and precise gene editing strategy. Nature Publishing Group UK 2020-10-23 /pmc/articles/PMC7584657/ /pubmed/33097693 http://dx.doi.org/10.1038/s41467-020-19136-7 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Schene, Imre F. Joore, Indi P. Oka, Rurika Mokry, Michal van Vugt, Anke H. M. van Boxtel, Ruben van der Doef, Hubert P. J. van der Laan, Luc J. W. Verstegen, Monique M. A. van Hasselt, Peter M. Nieuwenhuis, Edward E. S. Fuchs, Sabine A. Prime editing for functional repair in patient-derived disease models |
title | Prime editing for functional repair in patient-derived disease models |
title_full | Prime editing for functional repair in patient-derived disease models |
title_fullStr | Prime editing for functional repair in patient-derived disease models |
title_full_unstemmed | Prime editing for functional repair in patient-derived disease models |
title_short | Prime editing for functional repair in patient-derived disease models |
title_sort | prime editing for functional repair in patient-derived disease models |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584657/ https://www.ncbi.nlm.nih.gov/pubmed/33097693 http://dx.doi.org/10.1038/s41467-020-19136-7 |
work_keys_str_mv | AT scheneimref primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT jooreindip primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT okarurika primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT mokrymichal primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT vanvugtankehm primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT vanboxtelruben primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT vanderdoefhubertpj primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT vanderlaanlucjw primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT verstegenmoniquema primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT vanhasseltpeterm primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT nieuwenhuisedwardes primeeditingforfunctionalrepairinpatientderiveddiseasemodels AT fuchssabinea primeeditingforfunctionalrepairinpatientderiveddiseasemodels |