Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
Nat Commun. 2017 May 12;8:15315. doi: 10.1038/ncomms15315.
Efforts to manipulate locus-specific histone acetylation to assess their causal role in gene expression and cellular and behavioural phenotypes have been impeded by a lack of experimental tools. The Cas9 nuclease has been adapted to target epigenomic modifications, but a detailed description of the parameters of such synthetic epigenome remodellers is still lacking. Here we describe a Cas9-based histone deacetylase (HDAC) and the design principles required to achieve locus-specific histone deacetylation. We assess its range of activity and specificity, and analyse target gene expression in two different cell types to investigate cellular context-dependent effects. Our findings demonstrate that the chromatin environment is an important element to consider when utilizing this synthetic HDAC.
人们曾试图通过操控组蛋白乙酰化的特定位置来评估其在基因表达和细胞及行为表现方面的因果作用，但由于缺乏实验工具，这一尝试受到了阻碍。Cas9 核酸酶已被用于靶向表观遗传修饰，但对于此类合成表观基因组重塑酶的参数仍缺乏详细描述。在这里，我们描述了一种基于 Cas9 的组蛋白去乙酰化酶（HDAC）以及实现组蛋白特异性去乙酰化所需的设计原则。我们评估了其活性和特异性范围，并在两种不同的细胞类型中分析了靶基因的表达，以研究细胞上下文相关的影响。我们的研究结果表明，在利用这种合成的 HDAC 时，染色质环境是一个需要考虑的重要因素。
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