A sustained, cytoplasmic transgene expression system delivered by cationic liposomes.
A plasmid containing the reporter gene, chloramphenicol acetyltransferase (CAT), driven by the bacteriophage T7 promoter was co-delivered with purified T7 RNA polymerase by the DC-chol cationic liposomes into human embryonic kidney 293 cells to obtain a transient (2 days) CAT gene expression. To prolong the expression, a T7 autogene which contains the T7 RNA polymerase gene driven by the T7 promoter was included in the transfection complex as a self-amplifying regeneration mechanism for the polymerase. High level CAT gene expression was observed up to 5 days after transfection. This strong and sustained expression system should be useful in gene transfer experiments.
Gao
X
X
Department of Pharmacology, University of Pittsburgh, School of Medicine, PA 15261.
Jaffurs
D
D
Robbins
P D
PD
Huang
L
L
D002460:Cell Line; D003593:Cytoplasm; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D008081:Liposomes; D014764:Viral Proteins
D008081:Liposomes; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases