A modified CUT&RUN-seq technique for qPCR analysis of chromatin-protein interactions

Abstract

Summary: Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) even with optimization may give low signal-to-background ratio and spatial resolution. Here, we adapted Cleavage Under Targets and Release Using Nuclease (CUT&RUN) (originally developed by the Henikoff group) to develop CUT&RUN-qPCR. By studying the recruitment of selected proteins (but amenable to other proteins), we find that CUT&RUN-qPCR is more sensitive and gives better spatial resolution than ChIP-qPCR.For complete details on the use and execution of this protocol, please refer to Skene et al. (2018) and Skene and Henikoff (2017). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell-based Assays
• Molecular Biology
• Chromatin immunoprecipitation (ChIP)