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Easy quantification of template-directed CRISPR/Cas9 editing

Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE met...

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Detalles Bibliográficos
Autores principales: Brinkman, Eva K, Kousholt, Arne N, Harmsen, Tim, Leemans, Christ, Chen, Tao, Jonkers, Jos, van Steensel, Bas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6007333/
https://www.ncbi.nlm.nih.gov/pubmed/29538768
http://dx.doi.org/10.1093/nar/gky164
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author Brinkman, Eva K
Kousholt, Arne N
Harmsen, Tim
Leemans, Christ
Chen, Tao
Jonkers, Jos
van Steensel, Bas
author_facet Brinkman, Eva K
Kousholt, Arne N
Harmsen, Tim
Leemans, Christ
Chen, Tao
Jonkers, Jos
van Steensel, Bas
author_sort Brinkman, Eva K
collection PubMed
description Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE method for quantification of templated editing events, including point mutations. The resulting TIDER method is a rapid, cheap and accessible tool for testing and optimization of template-directed genome editing strategies. A free web tool for TIDER data analysis is available at http://tide.nki.nl.
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spelling pubmed-60073332018-06-25 Easy quantification of template-directed CRISPR/Cas9 editing Brinkman, Eva K Kousholt, Arne N Harmsen, Tim Leemans, Christ Chen, Tao Jonkers, Jos van Steensel, Bas Nucleic Acids Res Methods Online Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE method for quantification of templated editing events, including point mutations. The resulting TIDER method is a rapid, cheap and accessible tool for testing and optimization of template-directed genome editing strategies. A free web tool for TIDER data analysis is available at http://tide.nki.nl. Oxford University Press 2018-06-01 2018-03-10 /pmc/articles/PMC6007333/ /pubmed/29538768 http://dx.doi.org/10.1093/nar/gky164 Text en © The Author(s) 2018. 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 Non-Commercial 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 Methods Online
Brinkman, Eva K
Kousholt, Arne N
Harmsen, Tim
Leemans, Christ
Chen, Tao
Jonkers, Jos
van Steensel, Bas
Easy quantification of template-directed CRISPR/Cas9 editing
title Easy quantification of template-directed CRISPR/Cas9 editing
title_full Easy quantification of template-directed CRISPR/Cas9 editing
title_fullStr Easy quantification of template-directed CRISPR/Cas9 editing
title_full_unstemmed Easy quantification of template-directed CRISPR/Cas9 editing
title_short Easy quantification of template-directed CRISPR/Cas9 editing
title_sort easy quantification of template-directed crispr/cas9 editing
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6007333/
https://www.ncbi.nlm.nih.gov/pubmed/29538768
http://dx.doi.org/10.1093/nar/gky164
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