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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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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

5,446 Views

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10:49 min

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September 26, 2019

DOI:

10.3791/59910-v

DOI:
10.3791/59910-v

•

10:49 min
September 26, 2019

•

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

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

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Modur, V., Singh, N., Muhammad, B. A Murine Cell Line Based Model of Chronic CDK9 Inhibition to Study Widespread Non-Genetic Transcriptional Elongation Defects (TEdeff) in Cancers. J. Vis. Exp. (151), e59910, doi:10.3791/59910 (2019).

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