DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers.
Amphipathic asymmetric dendrimers have been investigated for use in delivery of genes into cells, with the objective of optimising transfection efficiency and maintaining cell viability. We have synthesised amphipathic asymmetric dendrimers by solid phase methods. The ability of two of these to transfect BHK cells in culture with beta-galactosidase gene was determined by X-gal staining. Cell viability was measured by the MTT assay for BHK cells, and by spectroscopy for lysis of erythrocytes. Interactions between dendrimer and DNA were investigated by agarose gel electrophoresis. BHK cells were optimally transfected at 5:1 +/- charge ratio yielding 20% cells receiving at least one copy of the plasmid. Cell viability decreased when the dendrimer to DNA ratio exceeded 5:1. Raising the pH significantly affected the electrophoretic mobility of complexes of dendrimer and DNA. We conclude that amphipathic asymmetric dendrimers enable efficient plasmid DNA uptake into BHK cells. Cell viability is maintained at high concentrations of dendrimer when complexed with DNA at a 5:1 +/- charge ratio. Efficiency of transfection and cell viability suggest the system may be suitable for gene delivery in vivo.
Shah
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DS
Department of Pharmaceutical and Biological Chemistry, The School of Pharmacy, University of London, 29/39, Brunswick Square, WC1N 1AX, London, UK.
Sakthivel
T
T
Toth
I
I
Florence
A T
AT
Wilderspin
A F
AF
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology*; D004847:Epithelial Cells / Q000187:drug effects* / Q000502:physiology; D004926:Escherichia coli; D005822:Genetic Vectors / Q000235:genetics / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D019814:Myristic Acid / Q000737:chemistry / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics / Q000494:pharmacology*; D011107:Polylysine / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011108:Polymers / Q000737:chemistry*; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics / Q000494:pharmacology*
D011108:Polymers; D019814:Myristic Acid; D011107:Polylysine; D001616:beta-Galactosidase