Burnett School of Biomedical Sciences, UCF College of Medicine, University of Central Florida, Orlando, FL USA.
Robert Wood Johnson Medical School Institute for Neurological Therapeutics, and Department of Neurology, Rutgers Biomedical and Health Sciences, Piscataway, NJ USA.
Epigenetics. 2022 Dec;17(13):2075-2081. doi: 10.1080/15592294.2022.2106646. Epub 2022 Aug 3.
Epigenetic regulation is a crucial factor controlling gene expression. Here, we report our CRISPR/dCas9-based modular epigenetic toolkit that enables gene-specific modulation of epigenetic architecture. By modifying the SunTag framework of dCas9 tagged with five GCN4 moieties, each epigenetic writer is bound to scFv and target-specific sgRNA, and this system is able to modify multiple epigenetic marks in a target-specific manner. We successfully demonstrated that this system is efficient in modifying individual histone post-translational modifications. We display its utility as a tool to understand the contributions of specific histone marks on gene expression by screening a large promoter region and identifying differential outcomes with high base-pair resolution. This epigenetic toolkit can be easily altered with a large variety of epigenetic effectors and is a useful tool for researchers to use in understanding gene-specific epigenetic changes and their relation to gene expression.
表观遗传调控是控制基因表达的关键因素。在这里，我们报告了基于 CRISPR/dCas9 的模块化表观遗传工具包，该工具包可实现基因特异性的表观遗传结构的调节。通过修饰带有五个 GCN4 结构域的 dCas9 的 SunTag 框架，每个表观遗传写入器都与 scFv 和靶向特异性 sgRNA 结合，该系统能够以靶向特异性方式修饰多个表观遗传标记。我们成功地证明了该系统在修饰单个组蛋白翻译后修饰方面非常有效。我们通过筛选大片段启动子区域并以高碱基分辨率识别差异结果，展示了其作为一种工具来了解特定组蛋白标记对基因表达的贡献的用途。该表观遗传工具包可以很容易地与多种表观遗传效应因子结合使用，是研究人员用于了解基因特异性表观遗传变化及其与基因表达关系的有用工具。
