Harnessing changes in open chromatin determined by ATAC-seq to generate insulin-responsive reporter constructs

Area of Science:

• Epigenetics
• Molecular Biology
• Gene Regulation

Background:

• Gene regulation is crucial for cellular function, with epigenomic studies uncovering complex transcriptional mechanisms.
• Assay for Transposase-Accessible Chromatin by sequencing (ATAC-seq) is a key epigenomic investigation tool.
• The potential for using ATAC-seq identified chromatin changes to create inducible reporter constructs remains largely unassayed.

Purpose of the Study:

• To investigate intervention-mediated changes in accessible chromatin using ATAC-seq.
• To determine if these changes can be used to generate intervention-inducible reporter constructs.
• To model these effects using the insulin signaling pathway in Drosophila S2 cells.

Main Methods:

• Performed ATAC-seq and RNA-seq on insulin-treated Drosophila S2 cells.
• Analyzed correlations between chromatin accessibility and gene expression data.
• Cloned candidate enhancer regions upstream of luciferase reporter genes.

Main Results:

• Found correlations between ATAC-seq and RNA-seq data, particularly for specific chromatin feature types.
• Identified a significant correlation between enhancer regions (1-2kb from TSS) and downstream gene expression.
• Demonstrated insulin-inducibility of cloned reporter constructs driven by identified enhancer regions.

Conclusions:

• Insulin-induced chromatin accessibility, identified via ATAC-seq, highlights functional enhancer regions.
• These enhancers can drive insulin-inducible reporter gene expression.
• ATAC-seq can identify regulatory elements for creating inducible gene expression systems.