TY - JOUR
T1 - Efficient production of HIV-1 viral-like particles in mouse cells
AU - Diaz-Griffero, Felipe
AU - Taube, Ran
AU - Muehlbauer, Stefan M.
AU - Brojatsch, Jürgen
N1 - Funding Information:
The authors thank R. Squires for critical reading of the manuscript, and D.R. Littman for the initial support of the project. R. Taube was supported by NIH Grants AI286 and AI49104-01, respectively. S.M. Muehlbauer and J. Brojatsch were supported by the NIH Medical Scientist Training Grant T32GM007288 and the USDA Grant 20043520415033, respectively.
PY - 2008/4/11
Y1 - 2008/4/11
N2 - Previous efforts to develop a mouse model for HIV/AIDS have been impaired by multiple blocks to HIV replication, including barriers to viral entry, proviral transcription, and assembly. Expression of human cofactors in murine cells overcomes early restrictions, but does not lead to the production of infectious HIV particles. Here we show that stable expression of a codon-optimized synthetic HIV-1 Gag-Pol construct (sGP) in murine cell lines results in efficient Gag production and viral-like particle (VLP) release. Stable expression of the sGP construct in murine cells such as NIH3T3 and A9 improved Gag processing resulting in efficient VLP release comparable to that found in human cells. Using highly efficient transient transfection procedures, we increased Gag expression, and were able to produce infectious HIV particles in NIH3T3 cells. However, the infectivity of VLPs produced in murine cells was significantly below that generated in 293T cells. Reduced infectivity of VLPs produced in murine cells correlated with lower HIV reporter RNA levels in these cells. Taken together, improving the expression of HIV-1 Gag-Pol by using the sGP construct overcomes, at least in part, late restrictions in murine cells.
AB - Previous efforts to develop a mouse model for HIV/AIDS have been impaired by multiple blocks to HIV replication, including barriers to viral entry, proviral transcription, and assembly. Expression of human cofactors in murine cells overcomes early restrictions, but does not lead to the production of infectious HIV particles. Here we show that stable expression of a codon-optimized synthetic HIV-1 Gag-Pol construct (sGP) in murine cell lines results in efficient Gag production and viral-like particle (VLP) release. Stable expression of the sGP construct in murine cells such as NIH3T3 and A9 improved Gag processing resulting in efficient VLP release comparable to that found in human cells. Using highly efficient transient transfection procedures, we increased Gag expression, and were able to produce infectious HIV particles in NIH3T3 cells. However, the infectivity of VLPs produced in murine cells was significantly below that generated in 293T cells. Reduced infectivity of VLPs produced in murine cells correlated with lower HIV reporter RNA levels in these cells. Taken together, improving the expression of HIV-1 Gag-Pol by using the sGP construct overcomes, at least in part, late restrictions in murine cells.
KW - Assembly
KW - Gag processing
KW - Human immunodeficiency virus
KW - Infectivity
KW - Maturation
KW - Mouse model
KW - Murine restrictions
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U2 - 10.1016/j.bbrc.2007.12.195
DO - 10.1016/j.bbrc.2007.12.195
M3 - Article
C2 - 18241668
AN - SCOPUS:39749164386
SN - 0006-291X
VL - 368
SP - 463
EP - 469
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -
