Using RNA-binding proteins for immunoprecipitation of mRNAs from Xenopus laevis embryos

Abstract

Summary: This protocol is developed for identifying mRNAs that form complexes with mRNA-binding proteins (mRBPs) in Xenopus laevis embryos at different developmental stages. Here, we describe the use of the Ybx1 mRBP for immunoprecipitation-based mRNA isolation. This protocol features the translation of the mRBP of interest directly in living embryos following injection of synthetic mRNA templates encoding a hybrid of this protein with a specific tag. This approach allows precipitation of mRNA-protein complexes from embryonic lysates using commercially available anti-tag antibodies.For complete details on the use and execution of this protocol, please refer to Parshina et al. (2020).

Keywords

• Model Organisms
• Molecular Biology
• Protein Biochemistry