A replication-deficient adenovirus enhances liposome-mediated nucleic acid transfer into a stable cell line expressing T7 RNA polymerase.
Liposome-mediated transfer of nucleic acids into a cell line expressing bacteriophage T7 RNA polymerase was enhanced by addition of a replication-deficient adenovirus (Ad5-259A) to transfection mixtures. Increasing quantities of Ad5-259A resulted in a dose-related (up to 30-fold) enhancement of reporter gene activity expressed in BT7-H cells transfected with plasmid DNA containing the reporter sequence fused to the internal ribosome entry site of encephalomyocarditis virus. Similarly, Ad5-259A enhanced reporter gene expression 7-fold following transfection of DNA containing the reporter sequence under transcriptional control of the Rous sarcoma virus long terminal repeat. Addition of Ad5-295A to transfection mixtures increased the proportion of cells staining positively for reporter gene activity, from 2 to 25% when the reporter was expressed via the T7 polymerase and from 20 to 50% when the reporter was under the control of a eucaryotic promoter. Thus, Ad5-259A enhanced reporter protein activities expressed by cytoplasmic T7-directed transcription and cap-independent initiation of translation, or nuclear transcription and cap-dependent translation. Transfection enhancement was blocked by neutralizing antibody to Ad5, and is most likely related to the endosome-disrupting activities of the virus. Adenovirus enhancement of liposome-mediated transfection provides a useful method for efficient nucleic acid transfer into eucaryotic cells.
Honda
M
M
Department of Medicine, University of North Carolina at Chapel Hill 27599-7030, USA.
Hu
P C
PC
Huang
C H
CH
Matsui
H
H
Lemon
S M
SM
D000260:Adenoviruses, Human / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D017123:Bacteriophage T7 / Q000201:enzymology; D002213:Capsid / Q000276:immunology; D036022:Capsid Proteins; D002460:Cell Line; D002461:Cell Line, Transformed; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D003673:Defective Viruses / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D016174:Hepacivirus / Q000235:genetics; D006801:Humans; D008081:Liposomes; D051379:Mice; D019251:Nucleocapsid / Q000235:genetics; D012313:RNA; D013194:Staining and Labeling; D014162:Transfection; D014779:Virus Replication
D000914:Antibodies, Viral; D036022:Capsid Proteins; D008081:Liposomes; C018315:hexon capsid protein, Adenovirus; D012313:RNA; D015500:Chloramphenicol O-Acetyltransferase; D012321:DNA-Directed RNA Polymerases