Positive selection system to screen for inhibitors of human immunodeficiency virus-1 transcription.
We describe a transcription-based assay system to screen for antiviral drugs in vivo. The system consists of two transcription units, a cytomegalovirus promoter driving a reporter gene physically linked to an HIV-1 promoter oriented in the opposite direction. Based on the arrangement of the transcription units, enhanced HIV-1 promoter activity in the presence of the viral transactivating Tat protein downregulates reporter gene expression initiated from the CMV promoter. Inhibitors of HIV-1 transcription relieve the suppression and are identified by an increase in reporter gene expression. This positive selection system allows discrimination between drugs that nonspecifically block cellular functions by cytotoxicity and molecules that specifically repress HIV-1 promoter activity.
Del Rosario
M
M
Chiron Technologies, Chiron Corporation, Emeryville, CA 94608, USA.
Stephans
J C
JC
Zakel
J
J
Escobedo
J
J
Giese
K
K
D000469:Alkaline Phosphatase / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology*; D003587:Cytomegalovirus / Q000235:genetics; D015536:Down-Regulation; D004353:Drug Evaluation, Preclinical; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D015696:Gene Products, tat / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000187:drug effects / Q000235:genetics; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*; D054322:tat Gene Products, Human Immunodeficiency Virus
D000998:Antiviral Agents; D015696:Gene Products, tat; D007527:Isoenzymes; D018894:Reverse Transcriptase Inhibitors; C055791:germ-cell AP isoenzyme; D054322:tat Gene Products, Human Immunodeficiency Virus; D054303:HIV Reverse Transcriptase; D000469:Alkaline Phosphatase