Effect of CRISPR-based epigenetic interference on cis-regulatory gene networks

• Main Author: Loh, Xinpeng
• Other Authors: Amartya Sanyal
• Format: Theses and Dissertations
• Language: English
• Published: 2018
• Subjects: DRNTU::Science::Biological sciences
• Online Access: https://hdl.handle.net/10356/80404; http://hdl.handle.net/10220/46576
• Institution: Nanyang Technological University

The revolutionary CRISPR-Cas9 genome engineering technology has been repurposed into a docking tool by fusing various effector proteins to the catalytically inactive Cas9. The modified system can regulate transcription by editing the epigenetic code, thereby modifying the local chromatin structure. While several reports have harnessed the potential of the CRISPR interference (CRISPRi) system to silence genes, only limited types of effector domains have been tested to date. Here, we expanded the repertoire of effector domains in the CRISPRi toolbox by incorporating novel epigenetic modulators - Histone Deacetylase 1 and Heterochromatin Protein 1 alpha - to study gene regulatory networks (GRNs). Efficiencies of these novel epigenetic effector domains were successfully demonstrated by targeting enhancers of globin genes in leukemia cells, as well as drug resistance genes in a chemoresistant model. Integration of new epigenetic modulators with CRISPRi would present new possibilities for understanding GRNs in various cell types and context.