Protocol to study the role of a human nuclear m6A RNA reader on chromatin-associated RNA targets

Abstract

Summary: Here, we present a detailed protocol to study the role of a human nuclear m6A RNA reader, YTHDC1, on chromatin-associated RNA targets. We describe steps for RNA extraction coupled to subnuclear fractionation to identify and study RNA-based regulations that take place in the chromatin-associated fraction. We then detail an RNA immunoprecipitation procedure adapted to identify chromatin-associated RNA targets. This protocol can be adapted to other human or mammalian chromatin-associated RNA binding proteins.For complete details on the use and execution of this protocol, please refer to Timcheva et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell separation/fractionation
• Gene Expression
• Molecular Biology
• Protein expression and purification