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.
Multiple epigenetic regulatory pathways collectively control gene expression and play key roles in various biological processes and human diseases. However, the functional relationships between different epigenetic mechanisms remain elusive. This proposal is to develop new enabling technologies to independently edit multiple epigenetic modifications that will allow the interrogation of crosstalk between distinct epigenetic pathways. We expect the results from this research will advance our understanding of gene regulation and lead to the development of new therapeutic strategies for human diseases.