Lineage-specific transgene expression in hematopoietic cells using a Cre-regulated retroviral vector

Vivian M Turner, Sandra Gardam, Robert Brink

Research output: Contribution to journalArticlepeer-review

Abstract

Transduction of bone marrow stem cells with retroviral expression vectors represents a cheaper and more rapid alternative to conventional transgenesis for studies of in vivo gene function. However, achieving tissue-specific expression of genes inserted into retroviral vectors is notoriously difficult. We have developed a single tri-cistronic retroviral vector (MG(f)I4) that facilitates Cre-dependent, lineage-specific gene expression within hematopoietic cells. Bone marrow stem cells transduced with MG(f)I4 co-express a loxP-flanked (floxed) eGFP cDNA together with truncated human CD4 (hCD4Delta). Open reading frames (ORFs) cloned between these two cDNAs are not constitutively translated but are activated upon Cre-mediated removal of the eGFP cDNA. Mice reconstituted with transduced bone marrow stem cells obtained from Cd19-Cre, Cr2-Cre or Lck-Cre, donors were shown to specifically express an ORF insert in the appropriate lymphocyte subsets. Cells that had activated ORF expression were identifiable by transition from a GFP+, hCD4+ to a GFP(-), hCD4+ phenotype. The use of this novel vector in conjunction with the wide range of well-characterized Cre-transgenic lines will be a versatile tool for exploring gene function within the immune system. In particular, this approach will provide a convenient way to test the functional significance of naturally occurring genetic mutations linked to human disease.

Original languageEnglish
Pages (from-to)162-6
Number of pages5
JournalJournal of Immunological Methods
Volume360
Issue number1-2
DOIs
Publication statusPublished - 31 Aug 2010
Externally publishedYes

Keywords

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Antigens, CD19
  • Antigens, CD4
  • Bone Marrow Cells
  • Bone Marrow Transplantation
  • Cell Lineage
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Hematopoietic Stem Cells
  • Humans
  • Integrases
  • Lymphocyte Subsets
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Organ Specificity
  • Receptors, Complement 3d
  • Retroviridae
  • Sequence Deletion

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