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

This protocol is developed for identifying mRNAs that form complexes with mRNA-binding proteins (mRBPs) in Xenopus laevis embryos at different develop-mental stages. Here, we describe the use of the Ybx1 mRBP for immunoprecipi-tation-based mRNA isolation. This protocol features the translation of the mRBP of interest directly in living embryos following injection of synthetic mRNA tem-plates 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).

Automated summary

This protocol is developed for identifying mRNAs that form complexes with mRNA-binding proteins in Xenopus laevis embryos at different developmental stages. It describes the use of the Ybx1 mRBP for immunoprecipitation-based mRNA isolation, featuring 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.