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...
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/5/4416 |
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| author | Wanxin Wang Peipei Huang Wenshuang Dai Huali Tang Yuliang Qiu Yanan Chang Zhiyang Han Xi Li Lipu Du Xingguo Ye Cheng Zou Ke Wang |
| author_facet | Wanxin Wang Peipei Huang Wenshuang Dai Huali Tang Yuliang Qiu Yanan Chang Zhiyang Han Xi Li Lipu Du Xingguo Ye Cheng Zou Ke Wang |
| author_sort | Wanxin Wang |
| collection | DOAJ |
| 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 <i>GUS</i> gene, the <i>Cas9</i> and a <i>GUS</i>-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 <i>GUS</i> 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 <i>TaWaxy</i> gene, which is responsible for amylose synthesis, was tested. Utilizing the aforementioned nicotinamide concentration to treat embryos containing the molecular components for editing the <i>TaWaxy</i> 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. |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d0131d4a150c4dc8a493a9ba01882e7b2023-11-17T07:48:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245441610.3390/ijms24054416Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid WheatWanxin Wang0Peipei Huang1Wenshuang Dai2Huali Tang3Yuliang Qiu4Yanan Chang5Zhiyang Han6Xi Li7Lipu Du8Xingguo Ye9Cheng Zou10Ke Wang11Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaNational Key Facility of Crop Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaInstitute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaHistone 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 <i>GUS</i> gene, the <i>Cas9</i> and a <i>GUS</i>-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 <i>GUS</i> 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 <i>TaWaxy</i> gene, which is responsible for amylose synthesis, was tested. Utilizing the aforementioned nicotinamide concentration to treat embryos containing the molecular components for editing the <i>TaWaxy</i> 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.https://www.mdpi.com/1422-0067/24/5/4416nicotinamidewheatCRISPR/Cas9histone deacetylase inhibitorsediting efficiencybase editing |
| spellingShingle | Wanxin Wang Peipei Huang Wenshuang Dai Huali Tang Yuliang Qiu Yanan Chang Zhiyang Han Xi Li Lipu Du Xingguo Ye Cheng Zou Ke Wang Application of Nicotinamide to Culture Medium Improves the Efficiency of Genome Editing in Hexaploid Wheat International Journal of Molecular Sciences nicotinamide wheat CRISPR/Cas9 histone deacetylase inhibitors editing efficiency base editing |
| 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 | nicotinamide wheat CRISPR/Cas9 histone deacetylase inhibitors editing efficiency base editing |
| url | https://www.mdpi.com/1422-0067/24/5/4416 |
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