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Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells
Precise gene editing in human pluripotent stem cells (hPSCs) holds great promise for studying and potentially treating human diseases. Both prime editing and base editing avoid introducing double strand breaks, but low editing efficiencies make those techniques still an arduous process in hPSCs. Her...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9613702/ https://www.ncbi.nlm.nih.gov/pubmed/36302757 http://dx.doi.org/10.1038/s41467-022-34045-7 |
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author | Li, Mu Zhong, Aaron Wu, Youjun Sidharta, Mega Beaury, Michael Zhao, Xiaolan Studer, Lorenz Zhou, Ting |
author_facet | Li, Mu Zhong, Aaron Wu, Youjun Sidharta, Mega Beaury, Michael Zhao, Xiaolan Studer, Lorenz Zhou, Ting |
author_sort | Li, Mu |
collection | PubMed |
description | Precise gene editing in human pluripotent stem cells (hPSCs) holds great promise for studying and potentially treating human diseases. Both prime editing and base editing avoid introducing double strand breaks, but low editing efficiencies make those techniques still an arduous process in hPSCs. Here we report that co-delivering of p53DD, a dominant negative fragment of p53, can greatly enhance prime editing and cytosine base editing efficiencies in generating precise mutations in hPSCs. We further apply PE3 in combination with p53DD to efficiently create multiple isogenic hPSC lines, including lines carrying GBA or LRRK2 mutations associated with Parkinson disease and a LMNA mutation linked to Hutchinson-Gilford progeria syndrome. We also correct GBA and LMNA mutations in the patient-specific iPSCs. Our data show that p53DD improves PE3 efficiency without compromising the genome-wide safety, making it feasible for safe and routine generation of isogenic hPSC lines for disease modeling. |
format | Online Article Text |
id | pubmed-9613702 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-96137022022-10-29 Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells Li, Mu Zhong, Aaron Wu, Youjun Sidharta, Mega Beaury, Michael Zhao, Xiaolan Studer, Lorenz Zhou, Ting Nat Commun Article Precise gene editing in human pluripotent stem cells (hPSCs) holds great promise for studying and potentially treating human diseases. Both prime editing and base editing avoid introducing double strand breaks, but low editing efficiencies make those techniques still an arduous process in hPSCs. Here we report that co-delivering of p53DD, a dominant negative fragment of p53, can greatly enhance prime editing and cytosine base editing efficiencies in generating precise mutations in hPSCs. We further apply PE3 in combination with p53DD to efficiently create multiple isogenic hPSC lines, including lines carrying GBA or LRRK2 mutations associated with Parkinson disease and a LMNA mutation linked to Hutchinson-Gilford progeria syndrome. We also correct GBA and LMNA mutations in the patient-specific iPSCs. Our data show that p53DD improves PE3 efficiency without compromising the genome-wide safety, making it feasible for safe and routine generation of isogenic hPSC lines for disease modeling. Nature Publishing Group UK 2022-10-27 /pmc/articles/PMC9613702/ /pubmed/36302757 http://dx.doi.org/10.1038/s41467-022-34045-7 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 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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Li, Mu Zhong, Aaron Wu, Youjun Sidharta, Mega Beaury, Michael Zhao, Xiaolan Studer, Lorenz Zhou, Ting Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title | Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title_full | Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title_fullStr | Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title_full_unstemmed | Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title_short | Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
title_sort | transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9613702/ https://www.ncbi.nlm.nih.gov/pubmed/36302757 http://dx.doi.org/10.1038/s41467-022-34045-7 |
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