STAR Protoc. 2022 May 25;3(2):101415. doi: 10.1016/j.xpro.2022.101415. eCollection 2022 Jun 17.

An RNA-immunoprecipitation protocol to identify RNAs associated with RNA-binding proteins in cytoplasmic and nuclear Drosophila head fractions

Marjorie Heim1, Lauren Blot1, Florence Besse1

1Université Côte d’Azur, CNRS, Inserm, Institut de Biologie Valrose, 06100 Nice, France.

RNA-binding proteins (RBPs) are multifunctional proteins that shuttle between the nucleus and the cytoplasm where they assemble with target RNAs to form multi-molecular complexes. Here, we describe a protocol to selectively identify RNAs associated with RBPs of interest in the cytoplasmic and nuclear compartments of adult Drosophila brain cells. Cytoplasmic and nuclear fractions are differentially collected and used for immunoprecipitation-based purification of GFP-tagged RBPs. This protocol can be applied to samples expressing ectopic or endogenous tagged RBPs.

DOI: 10.1016/j.xpro.2022.101415