Journal
STAR PROTOCOLS
Volume 2, Issue 2, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2021.100552
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Funding
- RFBR [20-34-90017, 18-04-00674 A]
- RSCF [20-34-70058]
- [19-14-00098]
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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.
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).
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