/
/
Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle
A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:04Title
- 01:24Preparation of S or M Phase-arrested Cells
- 02:19Formaldehyde Cross-linking
- 04:49Immunoprecipitation
- 07:32Results: Capturing and Analyzing PKR-interacting RNAs During the Cell Cycle Using qRT-PCR
- 08:45Conclusion
We present experimental approaches for studying RNA-interactors of double-stranded RNA binding protein kinase RNA-activated (PKR) during the mammalian cell cycle using HeLa cells. This method utilizes formaldehyde to crosslink RNA-PKR complexes and immunoprecipitation to enrich PKR-bound RNAs. These RNAs can be further analyzed through high-throughput sequencing or qRT-PCR.
Tags
Keywords:
RNA InteractorsProtein Kinase RNA-ActivatedMammalian Cell CycleCrosslinkingFormaldehydeImmunoprecipitationNeurodegenerative DiseaseParkinson’s DiseaseAlzheimer’s DiseaseDouble-stranded RNARNA-binding ProteinsPost-transcriptional RegulationGene ExpressionCell Cycle ArrestHeLa CellsThymidineNocodazole
