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Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques
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JoVE 杂志 免疫与感染
Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques

Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques

DOI:
10.3791/57776-v

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13:13 min

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September 25, 2018

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, , , ,
1US Military HIV Research Program, Henry M. Jackson Foundation,Walter Reed Army Institute of Research, 2Vaccine Research Center, NIAID,NIH

Chapters

  • 00:04Title
  • 00:46Gene Expression Assay Preparation and Surface Staining of Viable Cells
  • 03:18Prepare Cell Collection Plates, Perform FACS Sort, and Generate cDNA
  • 05:17Multiplex qPCR on the Biomark Platform
  • 11:26Results: Single-cell Quantitation of mRNA and Surface Protein Expression
  • 12:56Conclusion

Summary

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Described is a methodology to quantitate the expression of 96 genes and 18 surface proteins by single cells ex vivo, allowing for the identification of differentially expressed genes and proteins in virus-infected cells relative to uninfected cells. We apply the approach to study SIV-infected CD4+ T cells isolated from rhesus macaques.

Tags

Single-cell QuantitationMRNASurface ProteinSimian Immunodeficiency VirusCD4+ T CellsRhesus MacaquesHost-pathogen InteractionsCell SortingFlow CytometryGene Expression AssaysSurface StainingCell SuspensionCell ViabilityAntibody CocktailCell Strainer

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