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

ABSTRACT: 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).

SUBMITTER: Martynova NY

PROVIDER: S-EPMC8233257 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Martynova Natalia Y NY Parshina Elena A EA Zaraisky Andrey G AG

STAR protocols 20210524 2

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-pro ...[more]

PMID: 34195668

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