RNA–protein interactions are vital for numerous cellular processes, including transcriptional regulation, RNA splicing, RNA transport, translation control, and RNA degradation. During viral infections, the interaction between viral RNA and host proteins is crucial for viral replication, immune evasion, and the overall viral life cycle. Thus, investigating these interactions is essential for understanding viral mechanisms. RNA immunoprecipitation (RIP) is a powerful method for studying RNA–protein interactions, particularly in the context of viral infections. This technique leverages specific antibodies to enrich RNA–protein complexes, followed by RNA extraction and analysis, allowing for the identification of RNA-binding proteins involved in viral recognition, replication, and immune responses. RIP enables a comprehensive exploration of the dynamic changes in RNA–protein interactions and provides valuable insights into the roles of RNA-binding proteins in viral infections. This chapter introduces the principles and procedures of RIP, with a focus on the application of RIP-quantitative PCR (qPCR) for studying interactions between viral RNA and host proteins.

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RNA-Binding Protein Immunoprecipitation 1 (RIP) Assay to Investigate the Interactions Between Viral RNA and Host Proteins

  • Xiao Shan,
  • Zemin Ji,
  • Zhe Wang,
  • Penghui Hu,
  • Chunfu Zheng,
  • Qiujing Yu

摘要

RNA–protein interactions are vital for numerous cellular processes, including transcriptional regulation, RNA splicing, RNA transport, translation control, and RNA degradation. During viral infections, the interaction between viral RNA and host proteins is crucial for viral replication, immune evasion, and the overall viral life cycle. Thus, investigating these interactions is essential for understanding viral mechanisms. RNA immunoprecipitation (RIP) is a powerful method for studying RNA–protein interactions, particularly in the context of viral infections. This technique leverages specific antibodies to enrich RNA–protein complexes, followed by RNA extraction and analysis, allowing for the identification of RNA-binding proteins involved in viral recognition, replication, and immune responses. RIP enables a comprehensive exploration of the dynamic changes in RNA–protein interactions and provides valuable insights into the roles of RNA-binding proteins in viral infections. This chapter introduces the principles and procedures of RIP, with a focus on the application of RIP-quantitative PCR (qPCR) for studying interactions between viral RNA and host proteins.