Epigenetic features drastically impact CRISPR–Cas9 efficacy in plants

编号 040036801

推送时间 20221107

研究领域 森林培育 

年份 2022 

类型 期刊 

语种 英语

标题 Epigenetic features drastically impact CRISPR–Cas9 efficacy in plants

来源期刊 Plant Physiology

期 第368期

发表时间 20220611

关键词 CRISPR–Cas9;  Epigenetic features;  DNA;  Arabidopsis thaliana;  high-resolution epigenomic resources;  transgenic plants;  DNA methylation; 

摘要 CRISPR–Cas9-mediated genome editing has been widely adopted for basic and applied biological research in eukaryotic systems. While many studies consider DNA sequences of CRISPR target sites as the primary determinant for CRISPR mutagenesis efficiency and mutation profiles, increasing evidence reveals the substantial role of chromatin context. Nonetheless, most prior studies are limited by the lack of sufficient epigenetic resources and/or by only transiently expressing CRISPR–Cas9 in a short time window. In this study, we leveraged the wealth of high-resolution epigenomic resources in Arabidopsis (Arabidopsis thaliana) to address the impact of chromatin features on CRISPR–Cas9 mutagenesis using stable transgenic plants. Our results indicated that DNA methylation and chromatin features could lead to substantial variations in mutagenesis efficiency by up to 250-fold. Low mutagenesis efficiencies were mostly associated with repressive heterochromatic features. This repressive effect appeared to persist through cell divisions but could be alleviated through substantial reduction of DNA methylation at CRISPR target sites. Moreover, specific chromatin features, such as H3K4me1, H3.3, and H3.1, appear to be associated with significant variation in CRISPR–Cas9 mutation profiles mediated by the non-homologous end joining repair pathway. Our findings provide strong evidence that specific chromatin features could have substantial and lasting impacts on both CRISPR–Cas9 mutagenesis efficiency and DNA double-strand break repair outcomes.

服务人员 孙小满

服务院士 尹伟伦

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