Enhancement of calcium phosphate-mediated transfection by inclusion of adenovirus in coprecipitates
- 1 April 1999
- journal article
- research article
- Published by Springer Nature in Gene Therapy
- Vol. 6 (4) , 676-682
- https://doi.org/10.1038/sj.gt.3300857
Abstract
Although coprecipitates of plasmid DNA and calcium phosphate (DNA:CaPi coprecipitates) were one of the first methods used for transfection of mammalian cells, they are inefficient for many cell types. Based on the recent finding that incorporating recombinant adenovirus in CaPi coprecipitates enhances expression of virus-encoded transgenes, we tested the hypothesis that including adenovirus in DNA:CaPi coprecipitates would increase the efficiency of transfection. We found that including adenovirus at the time of coprecipitate formation increased transgene expression in several cell types. Only a short incubation with cells was required, and the coprecipitates could be delivered in the presence of serum. Inclusion of adenovirus in coprecipitates did not increase DNA uptake by cells, and inactivated virus was also effective. Neutralizing anti-hexon antibody attenuated the enhancement produced by incorporating virus. These data suggest that the virus enhanced expression at a step after cellular uptake, probably by increasing DNA release from endosomes. The DNA:CaPi:Ad coprecipitates were at least as effective as complexes of DNA with adenovirus and polethylenimine or Lipofectin, but produced less cellular toxicity. The results suggest that DNA:CaPi:Ad coprecipitates have advantages for in vitro gene transfer and provide an attractive vehicle for investigating cellular mechanisms of gene transfer.Keywords
This publication has 29 references indexed in Scilit:
- A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine.Proceedings of the National Academy of Sciences, 1995
- Gene transfer in vivo: sustained expression and regulation of genes introduced into the liver by receptor-targeted uptake.Proceedings of the National Academy of Sciences, 1994
- Hepatic gene therapy: adenovirus enhancement of receptor-mediated gene delivery and expression in primary hepatocytes.Proceedings of the National Academy of Sciences, 1993
- [41] Gene transfer into primary and established mammalian cell lines with lipopolyamine-coated DNAPublished by Elsevier ,1993
- Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes.Proceedings of the National Academy of Sciences, 1992
- High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles.Proceedings of the National Academy of Sciences, 1992
- High-Efficiency Gene Transfer Mediated by Adenovirus Coupled to DNA–Polylysine ComplexesHuman Gene Therapy, 1992
- A novel cationic liposome reagent for efficient transfection of mammalian cellsBiochemical and Biophysical Research Communications, 1991
- Direct Gene Transfer into Mouse Muscle in VivoScience, 1990
- Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure.Proceedings of the National Academy of Sciences, 1987