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Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat
Histone acetylation is the earliest and most well-characterized of post-translation modifications. It is mediated by histone acetyltransferases (HAT) and histone deacetylases (HDAC). Histone acetylation could change the chromatin structure and status and further regulate gene transcription. In this...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10002385/ https://www.ncbi.nlm.nih.gov/pubmed/36901844 http://dx.doi.org/10.3390/ijms24054416 |
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author | Wang, Wanxin Huang, Peipei Dai, Wenshuang Tang, Huali Qiu, Yuliang Chang, Yanan Han, Zhiyang Li, Xi Du, Lipu Ye, Xingguo Zou, Cheng Wang, Ke |
author_facet | Wang, Wanxin Huang, Peipei Dai, Wenshuang Tang, Huali Qiu, Yuliang Chang, Yanan Han, Zhiyang Li, Xi Du, Lipu Ye, Xingguo Zou, Cheng Wang, Ke |
author_sort | Wang, Wanxin |
collection | PubMed |
description | Histone acetylation is the earliest and most well-characterized of post-translation modifications. It is mediated by histone acetyltransferases (HAT) and histone deacetylases (HDAC). Histone acetylation could change the chromatin structure and status and further regulate gene transcription. In this study, nicotinamide, a histone deacetylase inhibitor (HDACi), was used to enhance the efficiency of gene editing in wheat. Transgenic immature and mature wheat embryos harboring a non-mutated GUS gene, the Cas9 and a GUS-targeting sgRNA were treated with nicotinamide in two concentrations (2.5 and 5 mM) for 2, 7, and 14 days in comparison with a no-treatment control. The nicotinamide treatment resulted in GUS mutations in up to 36% of regenerated plants, whereas no mutants were obtained from the non-treated embryos. The highest efficiency was achieved when treated with 2.5 mM nicotinamide for 14 days. To further validate the impact of nicotinamide treatment on the effectiveness of genome editing, the endogenous TaWaxy gene, which is responsible for amylose synthesis, was tested. Utilizing the aforementioned nicotinamide concentration to treat embryos containing the molecular components for editing the TaWaxy gene, the editing efficiency could be increased to 30.3% and 13.3%, respectively, for immature and mature embryos in comparison to the 0% efficiency observed in the control group. In addition, nicotinamide treatment during transformation progress could also improve the efficiency of genome editing approximately threefold in a base editing experiment. Nicotinamide, as a novel approach, may be employed to improve the editing efficacy of low-efficiency genome editing tools such as base editing and prime editing (PE) systems in wheat. |
format | Online Article Text |
id | pubmed-10002385 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-100023852023-03-11 Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat Wang, Wanxin Huang, Peipei Dai, Wenshuang Tang, Huali Qiu, Yuliang Chang, Yanan Han, Zhiyang Li, Xi Du, Lipu Ye, Xingguo Zou, Cheng Wang, Ke Int J Mol Sci Article Histone acetylation is the earliest and most well-characterized of post-translation modifications. It is mediated by histone acetyltransferases (HAT) and histone deacetylases (HDAC). Histone acetylation could change the chromatin structure and status and further regulate gene transcription. In this study, nicotinamide, a histone deacetylase inhibitor (HDACi), was used to enhance the efficiency of gene editing in wheat. Transgenic immature and mature wheat embryos harboring a non-mutated GUS gene, the Cas9 and a GUS-targeting sgRNA were treated with nicotinamide in two concentrations (2.5 and 5 mM) for 2, 7, and 14 days in comparison with a no-treatment control. The nicotinamide treatment resulted in GUS mutations in up to 36% of regenerated plants, whereas no mutants were obtained from the non-treated embryos. The highest efficiency was achieved when treated with 2.5 mM nicotinamide for 14 days. To further validate the impact of nicotinamide treatment on the effectiveness of genome editing, the endogenous TaWaxy gene, which is responsible for amylose synthesis, was tested. Utilizing the aforementioned nicotinamide concentration to treat embryos containing the molecular components for editing the TaWaxy gene, the editing efficiency could be increased to 30.3% and 13.3%, respectively, for immature and mature embryos in comparison to the 0% efficiency observed in the control group. In addition, nicotinamide treatment during transformation progress could also improve the efficiency of genome editing approximately threefold in a base editing experiment. Nicotinamide, as a novel approach, may be employed to improve the editing efficacy of low-efficiency genome editing tools such as base editing and prime editing (PE) systems in wheat. MDPI 2023-02-23 /pmc/articles/PMC10002385/ /pubmed/36901844 http://dx.doi.org/10.3390/ijms24054416 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wang, Wanxin Huang, Peipei Dai, Wenshuang Tang, Huali Qiu, Yuliang Chang, Yanan Han, Zhiyang Li, Xi Du, Lipu Ye, Xingguo Zou, Cheng Wang, Ke Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title | Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title_full | Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title_fullStr | Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title_full_unstemmed | Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title_short | Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat |
title_sort | application of nicotinamide to culture medium improves the efficiency of genome editing in hexaploid wheat |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10002385/ https://www.ncbi.nlm.nih.gov/pubmed/36901844 http://dx.doi.org/10.3390/ijms24054416 |
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