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Vpx mediated degradation of SAMHD1 has only a very limited effect on lentiviral transduction rate in ex vivo cultured HSPCs.
Journal article

Vpx mediated degradation of SAMHD1 has only a very limited effect on lentiviral transduction rate in ex vivo cultured HSPCs.

  • Li D Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland. Electronic address: duo.li@usz.ch.
  • Schlaepfer E Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland.
  • Audigé A Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland.
  • Rochat MA Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland.
  • Ivic S Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland.
  • Knowlton CN Center for Drug Discovery, Department of Pediatrics, Emory University, Atlanta, GA 30322, USA.
  • Kim B Center for Drug Discovery, Department of Pediatrics, Emory University, Atlanta, GA 30322, USA.
  • Keppler OT Institute of Medical Virology, National Reference Center for Retroviruses, University of Frankfurt, Frankfurt am Main 60054, Germany.
  • Speck RF Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Zurich, University of Zurich, 8091, Switzerland. Electronic address: Roberto.speck@usz.ch.
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  • 2015-07-25
Published in:
  • Stem cell research. - 2015
Gene engineering
Hematopoietic stem and progenitor cells (HSPCs)
Lentiviral vectors
SAMHD1
Transduction
dNTP pools
Cells, Cultured
Cytokines
HIV-1
Hematopoietic Stem Cells
Humans
Lentivirus
Monomeric GTP-Binding Proteins
RNA Interference
RNA, Small Interfering
SAM Domain and HD Domain-Containing Protein 1
Viral Regulatory and Accessory Proteins
English Understanding how to achieve efficient transduction of hematopoietic stem and progenitor cells (HSPCs), while preserving their long-term ability to self-reproduce, is key for applying lentiviral-based gene engineering methods. SAMHD1 is an HIV-1 restriction factor in myeloid and resting CD4+ T cells that interferes with reverse transcription by decreasing the nucleotide pools or by its RNase activity. Here we show that SAMHD1 is expressed at high levels in HSPCs cultured in a medium enriched with cytokines. Thus, we hypothesized that degrading SAMHD1 in HSPCs would result in more efficient lentiviral transduction rates. We used viral like particles (VLPs) containing Vpx, shRNA against SAMHD1, or provided an excess of dNTPs or dNs to study this question. Regardless of the method applied, we saw no increase in the lentiviral transduction rate. The result was different when we used viruses (HR-GFP-Vpx+) which carry Vpx and encode GFP. These viruses allow assessment of the effects of Vpx specifically in the transduced cells. Using HR-GFP-Vpx+ viruses, we observed a modest but significant increase in the transduction efficiency. These data suggest that SAMHD1 has some limited efficacy in blocking reverse transcription but the major barrier for efficient lentiviral transduction occurs before reverse transcription.
Language
  • English
Open access status
gold
Identifiers
Persistent URL
https://sonar.ch/global/documents/9630
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