Rivista
/
Genetica
/
Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification
JoVE Journal
Genetica
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Genetica
Transcriptome-Wide Profiling of Protein-RNA Interactions by Cross-Linking and Immunoprecipitation Mediated by FLAG-Biotin Tandem Purification

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

DOI:
10.3791/60730-v

•

08:46 min

•

May 18, 2020

•

, ,
1Tsinghua-Peking Center for Life Sciences, School of Medicine and School of Life Sciences,Tsinghua University, 2Department of Molecular Biology,University of Texas Southwestern Medical Center

Capitoli

  • 00:04Introduction
  • 00:41Crosslinking
  • 02:44Immunoprecipitation
  • 05:31Dephosphorylation and PNK Treatment
  • 06:53Results: WDR43 and LIN28 FbioCLIP-seq
  • 08:17Conclusion

Summary

Traduzione automatica

Traduzione automatica

Here we present a modified CLIP-seq protocol called FbioCLIP-seq with FLAG-biotin tandem purification to determine the RNA targets of RNA-binding proteins (RBPs) in mammalian cells.

Tags

Protein-RNA InteractionsCLIP-seqFLAG-BiotinTranscriptome-wide ProfilingRBPsMammalian CellsDirect Protein-bound RNAsIsotope-freeSDS-PAGEMembrane TransferMouse Embryonic Stem CellsUV Cross-linkingCell LysisDNase I TreatmentFLAG BeadsFLAG Peptide Elution

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

Analysis of the c-KIT Ligand Promoter Using Chromatin Immunoprecipitation

Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation

Promoter Capture Hi-C: High-resolution, Genome-wide Profiling of Promoter Interactions

Chromatin Immunoprecipitation of Murine Brown Adipose Tissue

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

An Assay for Quantifying Protein-RNA Binding in Bacteria

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

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii

Inducing Hairy Roots by Agrobacterium rhizogenes-Mediated Transformation in Tartary Buckwheat (Fagopyrum tataricum)

Read Article