Format

Send to

Choose Destination
Sci Rep. 2018 Oct 11;8(1):15036. doi: 10.1038/s41598-018-33042-5.

Robust Enhancement of Lentivirus Production by Promoter Activation.

Author information

1
Department of Molecular Virology, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
2
Department of Molecular Virology, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. takeshi-yoshida@umin.ac.jp.
3
Medical Affairs Unit, ViiV Healthcare K.K., 1-8-1 Akasaka, Minato-ku, Tokyo, 107-0052, Japan.
4
Department of Infectious Diseases and Immunology, Clinical Research Center, National Hospital Organization Nagoya Medical Center, Nagoya, Japan.
5
Department of Molecular Virology, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. shojmmb@tmd.ac.jp.

Abstract

Lentiviral vectors are a valuable tool to deliver exogenous genes for stable expression in cells. While much progress has been made in processing lentiviral vector-containing culture medium, it remains to be explored how the production of lentiviral vector from producer cells can be increased. We initially found that co-expression of the SPRY domain-containing SOCS box protein 1 (SPSB1) promotes the production of human immunodeficiency virus type 1 (HIV-1) and lentiviral vector with increased expression of the Gag and envelope proteins and activation of the HIV-1 LTR and CMV promoter. The presence of AP-1, NF-κB and CREB/ATF recognition sites in these promoters prompted us to utilize human T-lymphotropic virus type 1 (HTLV-1) Tax for lentiviral vector production because Tax activates all these transcription factors. Co-expression of a small amount of Tax markedly increased both the expression of viral structural proteins in producer cells and release of lentiviral vector particles, resulting in a more than 10-fold enhancement of transduction efficiency. Of note, the Tax protein was not detected in the lentiviral vector particles concentrated by ultracentrifugation, supporting the safety of this preparation. Collectively, these results indicate that promoter activation in producer cells represents a promising approach to preparing high-titer lentiviral vectors.

PMID:
30310119
PMCID:
PMC6181906
DOI:
10.1038/s41598-018-33042-5
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center