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Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo
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Biochemistry
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JoVE Journal Biochemistry
Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo

Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo

DOI:
10.3791/59624-v

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08:53 min

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May 25, 2019

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, , , , , ,
1Blood Research Institute,Blood Center of Wisconsin, 2Department of Medicine,Medical College of Wisconsin, 3Department of Surgery,The University of Alabama at Birmingham, 4Department of Biomedical Engineering,The University of Alabama at Birmingham, 5GBS Program, Graduate School,The University of Alabama at Birmingham

Chapters

  • 00:00Title
  • 02:34Methods
  • 07:06Results
  • 07:39Conclusion

Summary

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We present an approach to purify ribosome-bound mRNA from vascular endothelial cells (ECs) directly in mouse brain, lung and heart tissues via EC-specific genetic tag of enhanced green fluorescence protein (EGFP)in ribosomes in combination with RNA purification.

Tags

RNA IsolationEndothelial CellsRibosome Affinity Purification (TRAP)Transgenic MiceGFP-tagged RibosomesTranscriptome AnalysisAngiogenesisEpigeneticsTranscription RegulationGenotypingRNA SequencingRT-qPCR

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