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Overcoming the inhibitory effect of serum on lipofection by increasing the charge ratio of cationic liposome to DNA
- Source :
- Gene Therapy. 4:950-960
- Publication Year :
- 1997
- Publisher :
- Springer Science and Business Media LLC, 1997.
-
Abstract
- Since cationic liposome was first developed as a lipofection reagent, a drawback has been noted in that the efficiency of lipofection decreases dramatically after addition of serum to the lipofection medium. This drawback hampers the application of cationic liposome for systematic delivery of genes. In the present studies, we found that the effect of serum on DC-chol liposome-mediated lipofection is dependent on the charge ratio of liposome to DNA. Serum inhibited lipofection activity of the lipoplex at low charge ratios, whereas it enhanced the lipofection activity at high charge ratios. This phenomenon was observed using DOTAP/DOPE but not lipofectamine. Measurement of cellular association of DNA showed that serum could reduce the binding of lipoplex to cells at all tested charge ratios, i.e. 0-10.6. Removal of negatively charged proteins from serum by DEAE Sephacel column abolished the inhibitory effect of serum on lipofection. The fraction contained only negatively charged serum proteins which strongly inhibited lipofection at low charge ratios but not at higher charge ratios. Furthermore, preincubation of serum with positively charged polylysine, which neutralized negatively charged serum proteins, eliminated the inhibitory effect of serum on lipofection. In summary, inactivation of cationic liposome by serum is due to negatively charged serum proteins and it can be overcome by increasing charge ratio of cationic liposome-DNA lipoplexes or by neutralizing the serum with polylysine.
- Subjects :
- Genetic Vectors
Melanoma, Experimental
Synthetic membrane
Biology
Transfection
Mice
chemistry.chemical_compound
Cations
Tumor Cells, Cultured
Genetics
Animals
Humans
Polylysine
Cationic liposome
Luciferases
Molecular Biology
Liposome
Genetic transfer
Blood Proteins
DNA
Genetic Therapy
Molecular biology
Blood proteins
Rats
Blood
chemistry
Lipofectamine
Liposomes
Molecular Medicine
Electrophoresis, Polyacrylamide Gel
Subjects
Details
- ISSN :
- 14765462 and 09697128
- Volume :
- 4
- Database :
- OpenAIRE
- Journal :
- Gene Therapy
- Accession number :
- edsair.doi.dedup.....ae767833b535a9aca4c3fb61c1d595f2