Journal
/
Genética
/
Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)
JoVE Revista
Genética
Se requiere una suscripción a JoVE para ver este contenido.  Inicie sesión o comience su prueba gratuita.
JoVE Revista Genética
Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)

DOI:
10.3791/59913-v

•

09:26 min

•

July 10, 2019

•

, ,
1Department of Molecular Genetics, Center for RNA Biology,The Ohio State University

Capítulos

  • 00:04Título
  • 01:40Cell Harvesting and Formaldehyde Treatment
  • 02:50FLAG Immunoprecipitation and RNase I Digestion
  • 04:09Affinity Elution and Magnetic Bead-antibody Conjugation
  • 05:14Second Immunoprecipitation
  • 06:12RNA Extraction and Analysis
  • 07:51Results: RNA and Protein Yields
  • 08:38Conclusion

Summary

Traducción Automática

Traducción Automática

Here, we present a protocol to enrich endogenous RNA binding sites or "footprints" of RNA:protein (RNP) complexes from mammalian cells. This approach involves two immunoprecipitations of RNP subunits and is therefore dubbed RNA immunoprecipitation in tandem (RIPiT).

Tags

RNA ImmunoprecipitationRIPiT-SeqRNA-protein ComplexesEpitope-tagged ProteinCRISPRImmunoprecipitationRNA ExtractionHEK293 CellsRNA Binding ProteinsRNA ProcessingDiseasesNeuropathiesCancer

Article

Embed

Añadir a la lista de reproducción

Usage Statistics

Videos relacionados

Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis

Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets

Identification of Circular RNAs using RNA Sequencing

Tomato Root Transformation Followed by Inoculation with Ralstonia Solanacearum for Straightforward Genetic Analysis of Bacterial Wilt Disease

Screening and Identification of RNA Silencing Suppressors from Secreted Effectors of Plant Pathogens

Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification

Discovering CsgD Regulatory Targets in Salmonella Biofilm Using Chromatin Immunoprecipitation and High-Throughput Sequencing (ChIP-seq)

Read Article