Chemical and Light-Inducible Multiplex Epigenome Editing

项目摘要 这项提议的目标是开发新的工具来独立地和时空地操纵 明确基因座上不同的表观遗传调控机制。表观遗传修饰与染色质 结构调节基因的活动。表观遗传途径的改变和表观遗传调节因子的突变 导致许多发育性疾病和癌症。DNA甲基化和组蛋白修饰 被证明在基因激活中起着重要作用。一些研究表明，这些表观遗传途径是 被精心安排来调节活性基因的表达。然而，目前的方法仅限于精确地解剖。 这些不同的表观遗传调控机制之间的功能关系。为了解决这一限制， 我们建议开发独特的化学和光诱导方法，整合dCas9/gRNA引导 使用化学诱导邻近(CIP)技术进行靶向，以单独或同时控制 不同层次的表观遗传调控(例如，H3K27乙酰化和DNA去甲基化)。我们将使用这个 研究它们的功能相互作用并在表观基因组中测试多重表观遗传编辑的新技术。之后 完成这项工作，我们希望建立独特的工具，将大大有助于研究 基因激活中的表观遗传调控，并为开发基于表观遗传的新疗法提供新的方向。

Project Summary The objective of this proposal is to develop new tools to independently and spatiotemporally manipulate distinct epigenetic regulatory mechanisms at defined gene loci. Epigenetic modifications and chromatin structures regulate gene activity. Alteration of epigenetic pathways and mutations in epigenetic regulators contribute to many developmental diseases and cancers. DNA methylation and histone modification have been shown to play important roles in gene activation. Several studies suggest that these epigenetic pathways are orchestrated to regulate activity-gene expression. However, current methods are limited to precisely dissect the functional relationship between these distinct epigenetic regulatory mechanisms. To address this limitation, we propose to develop unique chemical and light inducible methods that integrate dCas9/gRNA-guided targeting with the chemically induced proximity (CIP) technology to individually or simultaneously control different layers of epigenetic regulations (e.g., H3K27 acetylation and DNA demethylation). We will use this new technology to study their functional interplays and test multiplex epigenetic editing in the epigenome. After finishing this work, we expect to establish unique tools that will significantly contribute to the studies of epigenetic regulation in gene activation and offer new directions in developing new epigenetic-based therapies.