RNA co-immunoprecipitation serves as a powerful technique for elucidating the interactions between RNA and RNA-binding proteins, pivotal for understanding posttranscriptional regulation mechanisms. This method captures the dynamics of protein–RNA associations across various cellular processes, applicable to both coding and noncoding RNAs. Here we describe a convenient and compelling method using nanobody coupled beads and UV-crosslinking. Importantly, this method can also be utilized to isolate short-lived complexes, for instance in RNA-degradation. The obtained RNA can be used for many downstream applications, such as qPCR or RNA-sequencing.

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Snapshot Moments of the Cell Revealed by UV-Crosslinking and RNA Co-immunoprecipitation of Transient RNA–Protein Complexes

  • Ivo Coban,
  • Heike Krebber

摘要

RNA co-immunoprecipitation serves as a powerful technique for elucidating the interactions between RNA and RNA-binding proteins, pivotal for understanding posttranscriptional regulation mechanisms. This method captures the dynamics of protein–RNA associations across various cellular processes, applicable to both coding and noncoding RNAs. Here we describe a convenient and compelling method using nanobody coupled beads and UV-crosslinking. Importantly, this method can also be utilized to isolate short-lived complexes, for instance in RNA-degradation. The obtained RNA can be used for many downstream applications, such as qPCR or RNA-sequencing.