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Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase
Histone phosphorylation is a ubiquitous post-translational modification that allows eukaryotic cells to rapidly respond to environmental stimuli. Despite correlative evidence linking histone phosphorylation to changes in gene expression, establishing the causal role of this key epigenomic modificati...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7873277/ https://www.ncbi.nlm.nih.gov/pubmed/33563994 http://dx.doi.org/10.1038/s41467-021-21188-2 |
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author | Li, Jing Mahata, Barun Escobar, Mario Goell, Jacob Wang, Kaiyuan Khemka, Pranav Hilton, Isaac B. |
author_facet | Li, Jing Mahata, Barun Escobar, Mario Goell, Jacob Wang, Kaiyuan Khemka, Pranav Hilton, Isaac B. |
author_sort | Li, Jing |
collection | PubMed |
description | Histone phosphorylation is a ubiquitous post-translational modification that allows eukaryotic cells to rapidly respond to environmental stimuli. Despite correlative evidence linking histone phosphorylation to changes in gene expression, establishing the causal role of this key epigenomic modification at diverse loci within native chromatin has been hampered by a lack of technologies enabling robust, locus-specific deposition of endogenous histone phosphorylation. To address this technological gap, here we build a programmable chromatin kinase, called dCas9-dMSK1, by directly fusing nuclease-null CRISPR/Cas9 to a hyperactive, truncated variant of the human MSK1 histone kinase. Targeting dCas9-dMSK1 to human promoters results in increased target histone phosphorylation and gene activation and demonstrates that hyperphosphorylation of histone H3 serine 28 (H3S28ph) in particular plays a causal role in the transactivation of human promoters. In addition, we uncover mediators of resistance to the BRAF V600E inhibitor PLX-4720 in human melanoma cells using genome-scale screening with dCas9-dMSK1. Collectively, our findings enable a facile way to reshape human chromatin using CRISPR/Cas9-based epigenome editing and further define the causal link between histone phosphorylation and human gene activation. |
format | Online Article Text |
id | pubmed-7873277 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-78732772021-02-24 Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase Li, Jing Mahata, Barun Escobar, Mario Goell, Jacob Wang, Kaiyuan Khemka, Pranav Hilton, Isaac B. Nat Commun Article Histone phosphorylation is a ubiquitous post-translational modification that allows eukaryotic cells to rapidly respond to environmental stimuli. Despite correlative evidence linking histone phosphorylation to changes in gene expression, establishing the causal role of this key epigenomic modification at diverse loci within native chromatin has been hampered by a lack of technologies enabling robust, locus-specific deposition of endogenous histone phosphorylation. To address this technological gap, here we build a programmable chromatin kinase, called dCas9-dMSK1, by directly fusing nuclease-null CRISPR/Cas9 to a hyperactive, truncated variant of the human MSK1 histone kinase. Targeting dCas9-dMSK1 to human promoters results in increased target histone phosphorylation and gene activation and demonstrates that hyperphosphorylation of histone H3 serine 28 (H3S28ph) in particular plays a causal role in the transactivation of human promoters. In addition, we uncover mediators of resistance to the BRAF V600E inhibitor PLX-4720 in human melanoma cells using genome-scale screening with dCas9-dMSK1. Collectively, our findings enable a facile way to reshape human chromatin using CRISPR/Cas9-based epigenome editing and further define the causal link between histone phosphorylation and human gene activation. Nature Publishing Group UK 2021-02-09 /pmc/articles/PMC7873277/ /pubmed/33563994 http://dx.doi.org/10.1038/s41467-021-21188-2 Text en © The Author(s) 2021 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 Li, Jing Mahata, Barun Escobar, Mario Goell, Jacob Wang, Kaiyuan Khemka, Pranav Hilton, Isaac B. Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title | Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title_full | Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title_fullStr | Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title_full_unstemmed | Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title_short | Programmable human histone phosphorylation and gene activation using a CRISPR/Cas9-based chromatin kinase |
title_sort | programmable human histone phosphorylation and gene activation using a crispr/cas9-based chromatin kinase |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7873277/ https://www.ncbi.nlm.nih.gov/pubmed/33563994 http://dx.doi.org/10.1038/s41467-021-21188-2 |
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