Enhanced adenovirus-assisted transfection composition and method

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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4352351, 536 231, C12N 1563, C12N 1511, C12N 700

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active

058307303

ABSTRACT:
A composition for transfecting eukaryotic cells comprising a cationic polymer which has protonatable groups which serve to buffer the acidic endosome, protecting the endocytosed polynucleotide from degradation and a viral agent is used to target uptake into and/or lysis from endosomes in the desired eukaryotic cell. By co-infecting the eukaryotic cells with cationic polymer, polynucleotide, and the viral agent, the polynucleotide is brought into the cell and then released. Preferably, polyamidoamine dendrimers are used as the cationic polymer and adenovirus is used as the viral agent. The dendrimers help associate plasmid DNA with the adenovirus, which then provokes receptor-mediated endocytosis. Within the endosome, the tertiary amine groups of the dendrimer buffer the pH change in the endosome. Then, the endosomalytic activity of the adenovirus releases the plasmid DNA into the cell. Also preferably, the adenovirus and the dendrimers are mixed before addition to the polynucleotide and the cells are washed after about 1.5 hours of incubation.

REFERENCES:
patent: 5661025 (1997-08-01), Szoka, Jr. et al.
Al Fasbender et al., "Complexes of Adenovirus with Polycationic Polymers and Cationic Lipids Increase the Efficiency of Gene Transfer in Vitro and in Vivo", The Journal of Biological Chemistry, vol. 272, No. 10, Issue of Mar. 7, 1997, pp. 6479-6489.

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