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

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10:49 min

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September 26, 2019

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

Chapters

  • 00:04Title
  • 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

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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.

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

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