Protocol for detecting genomic insulators in Drosophila using insulator-seq, a massively parallel reporter assay

Anastasiia Tonelli ¹, Pascal Cousin ², Maria Cristina Gambetta ³

 2024 Oct 24;5(4):103391.

• PMID: 39453817
• DOI: 10.1016/j.xpro.2024.103391

Free article

Abstract

Genomic insulators are DNA elements that prevent transcriptional activation of a promoter by an enhancer when interposed. We present a protocol for insulator-seq that enables high-throughput screening of genomic insulators using a plasmid-based massively parallel reporter assay in Drosophila cultured cells. We describe steps for insulator reporter plasmid library generation, transient transfection into cultured cells, and sequencing library preparation and provide a pipeline for data analysis. For complete details on the use and execution of this protocol, please refer to Tonelli et al.¹.