Journal
/
Genetik
/
A Murine Cell Line Based Model of Chronic CDK9 Inhibition to Study Widespread Non-Genetic Transcriptional Elongation Defects (TEdeff) in Cancers
JoVE Journal
Genetik
Zum Anzeigen dieser Inhalte ist ein JoVE-Abonnement erforderlich.  Melden Sie sich an oder starten Sie Ihre kostenlose Testversion.
JoVE Journal Genetik
A Murine Cell Line Based Model of Chronic CDK9 Inhibition to Study Widespread Non-Genetic Transcriptional Elongation Defects (TEdeff) in Cancers

A Murine Cell Line Based Model of Chronic CDK9 Inhibition to Study Widespread Non-Genetic Transcriptional Elongation Defects (TEdeff) in Cancers

DOI:
10.3791/59910-v

•

10:49 min

•

September 26, 2019

•

, ,
1Division of Experimental Hematology and Cancer Biology, Cancer and Blood Diseases Institute,Cincinnati Children’s Hospital Medical Center (CCHMC)

Kapitel

  • 00:04Titel
  • 01:28Confirmatory Assay of mRNA Processing Defects in the Generated Mouse TEdeff
  • 05:39Exploratory Assay of the Response of Mouse TEdeff Model to Antigen Specific Cytotoxic T-cell Attack
  • 08:14Results: Confirmatory and Exploratory Assessment of TEdeff Cell Model
  • 09:34Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

The protocol details an in vitro murine carcinoma model of non-genetic defective transcription elongation. Here, chronic inhibition of CDK9 is used to repress productive elongation of RNA Pol II along pro-inflammatory response genes to mimic and study the clinically observed TEdeff phenomenon, present in about 20% of all cancer types.

Tags

CDK9 InhibitionTranscriptional Elongation DefectsTEdeff CancersImmunotherapyPharmacological ModelMurine Cell LineIn VitroIn VivoT47DCAL51FlavopiridolPolyA-positive RNAOligo DT Magnetic BeadsRibosomal RNA DepletionBinding BufferWashing BufferTris-HCl

Article

Embed

ZUR WIEDERGABELISTE HINZUFÜGEN

Usage Statistics

Verwandte Videos

Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients

Laser Microirradiation to Study In Vivo Cellular Responses to Simple and Complex DNA Damage

The Nematode Caenorhabditis Elegans – A Versatile In Vivo Model to Study Host-microbe Interactions

Profiling DNA Replication Timing Using Zebrafish as an In Vivo Model System

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria

A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues

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

Navigating MARRVEL, a Web-Based Tool that Integrates Human Genomics and Model Organism Genetics Information

Read Article