Inhibition of HIV-1 replication by combined expression of gag dominant negative mutant and a human ribonuclease in a tightly controlled HIV-1 inducible vector.
An HIV-1-based expression vector has been constructed that produces protective genes tightly regulated by HIV-1 Tat and Rev proteins. The vector contains either a single protective gene (HIV-1 gag dominant negative mutant (delta-gag)) or a combination of two different protective genes (delta-gag and eosinophil-derived neurotoxin (EDN), a human ribonuclease) which are expressed from a dicistronic mRNA. After stable transfection of CEM T cells and following challenge with HIV-1, viral production was completely inhibited in cells transduced with the vector producing both delta-gag and EDN and delayed in cells producing delta-gag alone. In addition, cotransfection of HeLa-Tat cells with an infectious HIV-1 molecular clone and either protective vector demonstrated that the HIV-1 packaging signals present in the constructs were functional and allowed the efficient assembly of the protective RNAs into HIV-1 virions, thus potentially transmitting protection to the HIV-1 target cells.
Cara
A
A
Basic Research Laboratory, NCI, NIH, Bethesda, MD, USA.
Rybak
S M
SM
Newton
D L
DL
Crowley
R
R
Rottschafer
S E
SE
Reitz
M S
MS
Gusella
G L
GL
D015152:Blotting, Northern; D047208:Eosinophil-Derived Neurotoxin; D005786:Gene Expression Regulation; D015683:Gene Products, gag; D015694:Gene Products, rev; D015696:Gene Products, tat; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009498:Neurotoxins / Q000235:genetics; D016133:Polymerase Chain Reaction; D012260:Ribonucleases; D013602:T-Lymphocytes, Cytotoxic; D014162:Transfection; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015694:Gene Products, rev; D015696:Gene Products, tat; D009498:Neurotoxins; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus; D047208:Eosinophil-Derived Neurotoxin; D012260:Ribonucleases