Codelivery to mammalian cells of a transcriptional factor with cis-acting element using cationic liposomes.
The human immunodeficiency virus-1 transactivator protein (tat) was codelivered efficiently with a reporter gene under the control of a tat-responsive DNA element using different formulations of cationic liposomes. Expression of a tat-responsive reporter gene was induced by incubating cells with a mixture of purified recombinant tat protein, reporter DNA, and liposomes. Different cell lines were tested successfully as targets for the codelivery. Tat was shown to trans-activate the codelivered virus promoter specifically in the cells tested. Codelivery of tat with DNA is a useful model for studying the function of trans-acting factors and their cis-acting DNA elements. The currently available methods such as foot-printing only reveal the binding, but not the functional consequence of the binding, of the factor with the element. In addition, this system may prove useful as a model for high level and regulated transgene expression in target cells.
Farhood
H
H
Department of Pharmacology, University of Pittsburgh, School of Medicine, Pennsylvania 15261, USA.
Gao
X
X
Barsoum
J
J
Huang
L
L
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000096:biosynthesis; D006224:Cricetinae; D017931:DNA Primers; D004337:Drug Carriers; D015696:Gene Products, tat / Q000008:administration & dosage* / Q000096:biosynthesis; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007668:Kidney; D008081:Liposomes; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010714:Phosphatidylethanolamines; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000032:analysis / Q000096:biosynthesis; D017384:Sequence Deletion; D015533:Transcriptional Activation; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D054322:tat Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D004337:Drug Carriers; D015696:Gene Products, tat; D008081:Liposomes; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; C020888:1,2-dielaidoylphosphatidylethanolamine; D015500:Chloramphenicol O-Acetyltransferase