Protocol for identifying Dicer as dsRNA binding and cleaving reagent in response to transfected dsRNA.

Abstract

Mammalian Dicer has been proved to be functional on double-stranded RNAs (dsRNAs) and involved in antiviral immunity or immune regulation. Here, we present a protocol for identifying Dicer as a dsRNA binding and cleaving factor to transfected dsRNA in cell lines, based on small RNA sequencing (RNA-seq) and dsRNA-immunoprecipitation (dsRNA-IP). We detail both experimental processes and analysis on small RNA-seq data. This protocol can be further applied to identify proteins that process dsRNA into small RNAs, like RNase L. For complete details on the use and execution of this protocol, please refer to Dai et al.¹.