CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing

Scarless genome editing of induced pluripotent stem cells (iPSCs) is crucial for the precise modeling of genetic disease. Here we present CRISPR Del/Rei, a two-step deletion-reinsertion strategy with high editing efficiency and simple PCR-based screening that generates isogenic clones in ~ 2 months....

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Autores principales: Feuer, Kyra L., Wahbeh, Marah H., Yovo, Christian, Rabie, Eman, Lam, Anh-Thu N., Abdollahi, Sara, Young, Lindsay J., Rike, Bailey, Umamageswaran, Akul, Avramopoulos, Dimitrios
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
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Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279498/
https://www.ncbi.nlm.nih.gov/pubmed/35831384
http://dx.doi.org/10.1038/s41598-022-16004-w
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author Feuer, Kyra L.
Wahbeh, Marah H.
Yovo, Christian
Rabie, Eman
Lam, Anh-Thu N.
Abdollahi, Sara
Young, Lindsay J.
Rike, Bailey
Umamageswaran, Akul
Avramopoulos, Dimitrios
author_facet Feuer, Kyra L.
Wahbeh, Marah H.
Yovo, Christian
Rabie, Eman
Lam, Anh-Thu N.
Abdollahi, Sara
Young, Lindsay J.
Rike, Bailey
Umamageswaran, Akul
Avramopoulos, Dimitrios
author_sort Feuer, Kyra L.
collection PubMed
description Scarless genome editing of induced pluripotent stem cells (iPSCs) is crucial for the precise modeling of genetic disease. Here we present CRISPR Del/Rei, a two-step deletion-reinsertion strategy with high editing efficiency and simple PCR-based screening that generates isogenic clones in ~ 2 months. We apply our strategy to edit iPSCs at 3 loci with only rare off target editing.
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spelling pubmed-92794982022-07-15 CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing Feuer, Kyra L. Wahbeh, Marah H. Yovo, Christian Rabie, Eman Lam, Anh-Thu N. Abdollahi, Sara Young, Lindsay J. Rike, Bailey Umamageswaran, Akul Avramopoulos, Dimitrios Sci Rep Article Scarless genome editing of induced pluripotent stem cells (iPSCs) is crucial for the precise modeling of genetic disease. Here we present CRISPR Del/Rei, a two-step deletion-reinsertion strategy with high editing efficiency and simple PCR-based screening that generates isogenic clones in ~ 2 months. We apply our strategy to edit iPSCs at 3 loci with only rare off target editing. Nature Publishing Group UK 2022-07-13 /pmc/articles/PMC9279498/ /pubmed/35831384 http://dx.doi.org/10.1038/s41598-022-16004-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Feuer, Kyra L.
Wahbeh, Marah H.
Yovo, Christian
Rabie, Eman
Lam, Anh-Thu N.
Abdollahi, Sara
Young, Lindsay J.
Rike, Bailey
Umamageswaran, Akul
Avramopoulos, Dimitrios
CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title_full CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title_fullStr CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title_full_unstemmed CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title_short CRISPR Del/Rei: a simple, flexible, and efficient pipeline for scarless genome editing
title_sort crispr del/rei: a simple, flexible, and efficient pipeline for scarless genome editing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9279498/
https://www.ncbi.nlm.nih.gov/pubmed/35831384
http://dx.doi.org/10.1038/s41598-022-16004-w
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