Specific marking of HIV-1 positive cells using a Rev-dependent lentiviral vector expressing the green fluorescent protein.
Most of HIV-responsive expression vectors are based on the HIV promoter, the long terminal repeat (LTR). While responsive to an early HIV protein, Tat, the LTR is also responsive to cellular activation states and to the local chromatin activity where the integration has occurred. This can result in high HIV-independent activity, and has restricted the usefulness of LTR-based reporter to mark HIV positive cells. Here, we constructed an expression lentiviral vector that possesses, in addition to the Tat-responsive LTR, numerous HIV DNA sequences that include the Rev-response element and HIV splicing sites. The vector was incorporated into a lentiviral reporter virus, permitting highly specific detection of replicating HIV in living cell populations. The activity of the vector was measured by expression of the green fluorescence protein (GFP). The application of this vector as reported here offers a novel alternative approach to existing methods, such as in situ PCR or HIV antigen staining, to identify HIV-positive cells. The vector can also express therapeutic genes for basic or clinical experimentation to target HIV-positive cells.
National Center for Biodefense and Infectious Diseases, Department of Molecular and Microbiology, George Mason University, USA.
Jia
J
Guo
Clinton
C
Enos
Yuntao
Y
Wu
D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D020218:Response Elements; D014162:Transfection / Q000379:methods; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1; D049452:Green Fluorescent Proteins