Targeting chromatin regulators to specific genomic locations for gene control is emerging as a powerful method in basic research and synthetic biology. However, many chromatin regulators are large, making them difficult to deliver and combine in mammalian cells. Here, we developed a new strategy for gene control using small nanobodies that bind and recruit endogenous chromatin regulators to a gene. We show that an antiGFP nanobody can be used to simultaneously visualize GFP-tagged chromatin regulators and control gene expression, and that nanobodies against HP1 and DNMT1 can silence a reporter gene. Moreover, combining nanobodies together or with other regulators, such as DNMT3A or KRAB, can enhance silencing speed and epigenetic memory. Finally, we use the slow silencing speed and high memory of antiDNMT1 to build a signal duration timer and recorder. These results set the basis for using nanobodies against chromatin regulators for controlling gene expression and epigenetic memory.

中文翻译：

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纳米抗体介导的基因表达和表观遗传记忆的控制

在基础研究和合成生物学中，将染色质调节剂靶向特定基因组位置进行基因控制正在成为一种强大的方法。然而，许多染色质调节剂很大，使其难以在哺乳动物细胞中传递和结合。在这里，我们开发了一种使用小型纳米抗体进行基因控制的新策略，这些纳米抗体将内源性染色质调节剂结合并募集到一个基因上。我们表明，antiGFP纳米抗体可用于同时可视化GFP标记的染色质调节剂和控制基因表达，并且针对HP1和DNMT1的纳米抗体可以使报告基因沉默。此外，将纳米抗体结合在一起或与其他调节剂（例如DNMT3A或KRAB）结合使用，可以提高沉默速度和表观遗传记忆。最后，我们使用antiDNMT1的低沉默速度和高内存来构建信号持续时间计时器和记录器。这些结果为使用抗染色质调节剂的纳米抗体控制基因表达和表观遗传记忆奠定了基础。