TY  - JOUR
A1  - Li, Duo
A1  - Schlaepfer, Erika
A1  - Audigé, Annette
A1  - Rochat, Mary-Aude
A1  - Ivic, Sandra
A1  - Knowlton, Caitlin N.
A1  - Kimb, Baek
A1  - Keppler, Oliver Till
A1  - Speck, Roberto F.
T1  - Vpx mediated degradation of SAMHD1 has only a very limited effect on lentiviral transduction rate in ex vivo cultured HSPCs
T2  - Stem cell research
N2  - 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.
KW  - Gene engineering
KW  - Hematopoietic stem and progenitor cells (HSPCs)
KW  - Lentiviral vectors
KW  - SAMHD1
KW  - Transduction
KW  - dNTP pools
Y1  - 2015
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/40342
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-403423
SN  - 1873-5061
SN  - 1876-7753
N1  - © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
VL  - 15
SP  - 271
EP  - 280
PB  - Elsevier
CY  - Amsterdam [u.a.]
ER  -