RNA APPROACH FOR GENE SPECIFIC DEMETHYLATION

LIU YANJING

Publication

RNA APPROACH FOR GENE SPECIFIC DEMETHYLATION

LIU YANJING

Abstract

Methylation of CpG-rich promoters is associated with long-term gene silencing in malignant cells, thus a therapeutic approach reverting this mechanism represents a strategy to restore expression of aberrantly methylated genes. Currently, low toxicity and gene-specific demethylating agents are still missing. In this thesis, we successfully developed CRISPR-DiR as a gene specific demethylation and activation tool, in which DNMT1-interacting RNAs (DiRs) loops are fused to CRISPR sgRNA and are delivered to a specific locus to block DNMT1. By designing guides targeting the tumor suppressor gene p16, we successfully demethylated p16 and restore gene expression. In addition, the successful application of CRISPR-DiR to SALL4 gene indicates that it can be a general approach for multiple genes. This thesis explored the possibility to use RNA as gene-specific demethylating tool, which leads to the understanding of the epigenetic regulation process and paves the way for development of RNA-based gene-specific demethylating tool for cancer treatment.