Adenovirus-mediated augmentation of cell transfection with unmodified plasmid vectors

Kunihiko Yoshimura, Melissa A. Rosenfeld, Prem Seth, Ronald Crystal

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

The present study demonstrates that the human adenovirus (Ad) can augment transfer and expression of a gene within plasmid DNA unmodified by nonspecific linkers or by linker-ligand complexes. Following the transfection of COS-7 cells with pRSVL, a luciferase expression plasmid vector directed by the Rous sarcoma virus-long terminal repeat promoter, luciferase activity in the target cells was 103- to 104-fold higher when the cells were also infected with Ad-CFTR, a replication-deficient recombinant Ad containing human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA. The enhancement of luciferase gene expression in COS-7 cells was also observed with Ad-dl312 (a replication-deficient E1a deletion mutant Ad with no exogenous gene) and wild type Ad5. The efficiency of cell transfection with pRSVL in the presence of an Ad was achieved in a dose-dependent fashion with progressively higher luciferase activity in cells infected by increasing amounts of Ad-CFTR, Ad-dl312, or Ad5. The augmentation by Ad-CFTR of the transfer and expression of the luciferase gene in cells was similar to that of another transfection reagent, cationic liposomes. Further, when Ad-CFTR and liposomes were used in combination, 4- to 100-fold more efficient expression of the luciferase gene was achieved than with Ad-CFTR or liposomes alone. When COS-7, HeLa, and CV-1 cells were evaluated in parallel in the presence or absence of liposomes, Ad-mediated enhancement of luciferase activity was observed in all cell lines. Thus, exposure of target cells to replication-deficient or competent human Ad will markedly augment transfer and expression of the genes within plasmid DNA in mammalian cells in vitro without modifying the plasmid with linkers or linker-ligand complexes, a strategy that should be useful for in vitro and in vivo gene transfer applications.

Original languageEnglish
Pages (from-to)2300-2303
Number of pages4
JournalJournal of Biological Chemistry
Volume268
Issue number4
Publication statusPublished - 5 Feb 1993
Externally publishedYes

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Cystic Fibrosis Transmembrane Conductance Regulator
Luciferases
Adenoviridae
Transfection
Plasmids
Liposomes
Genes
Human Adenoviruses
Gene Expression
COS Cells
Cells
Ligands
Gene transfer
Terminal Repeat Sequences
DNA
Viruses
Gene expression
Rous sarcoma virus
Complementary DNA
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Adenovirus-mediated augmentation of cell transfection with unmodified plasmid vectors. / Yoshimura, Kunihiko; Rosenfeld, Melissa A.; Seth, Prem; Crystal, Ronald.

In: Journal of Biological Chemistry, Vol. 268, No. 4, 05.02.1993, p. 2300-2303.

Research output: Contribution to journalArticle

Yoshimura, Kunihiko ; Rosenfeld, Melissa A. ; Seth, Prem ; Crystal, Ronald. / Adenovirus-mediated augmentation of cell transfection with unmodified plasmid vectors. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 4. pp. 2300-2303.
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