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Efficient genome editing of differentiated renal epithelial cells
Recent advances in genome editing technologies have enabled the rapid and precise manipulation of genomes, including the targeted introduction, alteration, and removal of genomic sequences. However, respective methods have been described mainly in non-differentiated or haploid cell types. Genome edi...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5222933/ https://www.ncbi.nlm.nih.gov/pubmed/27987038 http://dx.doi.org/10.1007/s00424-016-1924-4 |
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author | Hofherr, Alexis Busch, Tilman Huber, Nora Nold, Andreas Bohn, Albert Viau, Amandine Bienaimé, Frank Kuehn, E. Wolfgang Arnold, Sebastian J. Köttgen, Michael |
author_facet | Hofherr, Alexis Busch, Tilman Huber, Nora Nold, Andreas Bohn, Albert Viau, Amandine Bienaimé, Frank Kuehn, E. Wolfgang Arnold, Sebastian J. Köttgen, Michael |
author_sort | Hofherr, Alexis |
collection | PubMed |
description | Recent advances in genome editing technologies have enabled the rapid and precise manipulation of genomes, including the targeted introduction, alteration, and removal of genomic sequences. However, respective methods have been described mainly in non-differentiated or haploid cell types. Genome editing of well-differentiated renal epithelial cells has been hampered by a range of technological issues, including optimal design, efficient expression of multiple genome editing constructs, attainable mutation rates, and best screening strategies. Here, we present an easily implementable workflow for the rapid generation of targeted heterozygous and homozygous genomic sequence alterations in renal cells using transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR) system. We demonstrate the versatility of established protocols by generating novel cellular models for studying autosomal dominant polycystic kidney disease (ADPKD). Furthermore, we show that cell culture-validated genetic modifications can be readily applied to mouse embryonic stem cells (mESCs) for the generation of corresponding mouse models. The described procedure for efficient genome editing can be applied to any cell type to study physiological and pathophysiological functions in the context of precisely engineered genotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00424-016-1924-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5222933 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-52229332017-01-19 Efficient genome editing of differentiated renal epithelial cells Hofherr, Alexis Busch, Tilman Huber, Nora Nold, Andreas Bohn, Albert Viau, Amandine Bienaimé, Frank Kuehn, E. Wolfgang Arnold, Sebastian J. Köttgen, Michael Pflugers Arch Molecular and Genomic Physiology Recent advances in genome editing technologies have enabled the rapid and precise manipulation of genomes, including the targeted introduction, alteration, and removal of genomic sequences. However, respective methods have been described mainly in non-differentiated or haploid cell types. Genome editing of well-differentiated renal epithelial cells has been hampered by a range of technological issues, including optimal design, efficient expression of multiple genome editing constructs, attainable mutation rates, and best screening strategies. Here, we present an easily implementable workflow for the rapid generation of targeted heterozygous and homozygous genomic sequence alterations in renal cells using transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR) system. We demonstrate the versatility of established protocols by generating novel cellular models for studying autosomal dominant polycystic kidney disease (ADPKD). Furthermore, we show that cell culture-validated genetic modifications can be readily applied to mouse embryonic stem cells (mESCs) for the generation of corresponding mouse models. The described procedure for efficient genome editing can be applied to any cell type to study physiological and pathophysiological functions in the context of precisely engineered genotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00424-016-1924-4) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2016-12-16 2017 /pmc/articles/PMC5222933/ /pubmed/27987038 http://dx.doi.org/10.1007/s00424-016-1924-4 Text en © The Author(s) 2016 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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. |
spellingShingle | Molecular and Genomic Physiology Hofherr, Alexis Busch, Tilman Huber, Nora Nold, Andreas Bohn, Albert Viau, Amandine Bienaimé, Frank Kuehn, E. Wolfgang Arnold, Sebastian J. Köttgen, Michael Efficient genome editing of differentiated renal epithelial cells |
title | Efficient genome editing of differentiated renal epithelial cells |
title_full | Efficient genome editing of differentiated renal epithelial cells |
title_fullStr | Efficient genome editing of differentiated renal epithelial cells |
title_full_unstemmed | Efficient genome editing of differentiated renal epithelial cells |
title_short | Efficient genome editing of differentiated renal epithelial cells |
title_sort | efficient genome editing of differentiated renal epithelial cells |
topic | Molecular and Genomic Physiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5222933/ https://www.ncbi.nlm.nih.gov/pubmed/27987038 http://dx.doi.org/10.1007/s00424-016-1924-4 |
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