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Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells
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JoVE 杂志 遗传学
Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells

Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells

DOI:
10.3791/60654-v

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10:49 min

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March 06, 2020

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, , , ,
1Centre de Recherche du Centre hospitalier de l’Université de Montréal (CRCHUM), 2Institut de Recherches Cliniques de Montréal (IRCM), 3Département de médecine, Faculté de médecine,Université de Montréal

Chapters

  • 00:05Introduction
  • 00:43Response Plasmid Design and Generation
  • 02:10Response Plasmid Transfection
  • 05:25Transcription Inhibition
  • 06:27mRNA Stability Analysis
  • 08:22Results: Representative mRNA Transcript Stability Evaluation
  • 10:20Conclusion

Summary

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Here, we present a tool that can be used to study the posttranscriptional modulation of a transcript in primary alveolar epithelial cells by using an inducible expression system coupled to a pipette electroporation technique.

Tags

Transcriptional ControlPlasmid Expression SystemMRNA StabilityPrimary Alveolar Epithelial CellsTransient TransfectionResponse PlasmidGene Of InterestV5 Epitope3′ UTRRestriction AnalysisSequencingPrimary Rat Lung Type II Alveolar Epithelial CellsCell CultureTransfection

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