INVESTIGATION OF ENHANCER–PROMOTER INTERACTIONS IN HUMAN DISEASE USING CRISPR-BASED FUNCTIONAL APPROACHES

Abstract

Interactions between Enhancer and promoters are vital in the process of controlling the expression rate of the genes since they allow distal regulatory factors to come in physical contact with target promoters of the genes within the three-dimensional chromatin scaffolding. The maladaptation of these interactions has been more and more associated with numerous human diseases, such as cancer, neurological and complex genetic disorders. Conventional experimental technologies like chromosome conformation capture (3C/Hi-C) and chromatin immunoprecipitation sequencing (ChIP-seq) have also been useful to understand genome organisation, but these techniques are mainly correlation based and do not provide causal relationships. New developments in CRISPR-based functional genomics have completely changed the landscape of gene regulation research, allowing specific and accurate perturbation of enhancer components. CRISPR-Cas9-mediated deletion, CRISPR interference (CRISPRi), and CRISPR activation (CRISPRa) can be used to obtain systematic studies about the role of enhancers and their role in disease-related gene expression. Also, regulatory networks can now be resolved and interpreted more deeply, with the combination of CRISPR screening and multi-omics data, such as RNA sequencing and chromatin accessibility profiling. The current review has given an overview of contemporary information on Enhancer–promoter interaction; emphasised CRISPR based methods in functional validation; and given a discussion on the new challenges and future patients on the understanding of mechanisms of gene regulation in human disease.