TSA promotes CRISPR/Cas9 editing efficiency and expression of cell division-related genes from plant protoplasts

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Title
TSA promotes CRISPR/Cas9 editing efficiency and expression of cell division-related genes from plant protoplasts
Author(s)
Seung Hee Choi; M H Lee; Da Mon Jin; Su Ji Ju; Woo-Seok Ahn; Eun Yee Jie; Ji Min Lee; Jiyoung LeeCha Young KimSuk Weon Kim
Bibliographic Citation
International Journal of Molecular Sciences, vol. 22, no. 15, pp. 7817-7817
Publication Year
2021
Abstract
Trichostatin A (TSA) is a representative histone deacetylase (HDAC) inhibitor that modulates epigenetic gene expression by regulation of chromatin remodeling in cells. To investigate whether the regulation of chromatin de-condensation by TSA can affect the increase in the efficiency of Cas9 protein-gRNA ribonucleoprotein (RNP) indel formation from plant cells, genome editing efficiency using lettuce and tobacco protoplasts was examined after several concentrations of TSA treatments (0, 0.1, 1 and 10 μM). RNP delivery from protoplasts was conducted by conventional polyethylene glycol (PEG) transfection protocols. Interestingly, the indel frequency of the SOC1 gene from TSA treatments was about 3.3 to 3.8 times higher than DMSO treatment in lettuce protoplasts. The TSA-mediated increase of indel frequency of the SOC1 gene in lettuce protoplasts occurred in a concentration-dependent manner, although there was not much difference. Similar to lettuce, TSA also increased the indel frequency by 1.5 to 1.8 times in a concentration-dependent manner during PDS genome editing using tobacco protoplasts. The MNase test clearly showed that chromatin accessibility with TSA treatments was higher than that of DMSO treatment. Additionally, TSA treatment significantly increased the level of histone H3 and H4 acetylation from lettuce protoplasts. The qRT-PCR analysis showed that expression of cell division-related genes (LsCYCD1-1, LsCYCD3-2, LsCYCD6-1, and LsCYCU4-1) was increased by TSA treatment. These findings could contribute to increasing the efficiency of CRISPR/Cas9-mediated genome editing. Furthermore, this could be applied for the development of useful genome-edited crops using the CRISPR/Cas9 system with plant protoplasts. ⓒ 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Keyword
Trichostatin AGenome editing efficiencyHistone acetylationChromatin de-condensationPlant protoplastsLettuceTobacco
ISSN
1661-6596
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/ijms22157817
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
Jeonbuk Branch Institute > 1. Journal Articles
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