Cell transfection with polycationic cyclodextrin vectors.
Polycationic cyclodextrins (CDs) were complexed with plasmid DNA and their effectiveness as vectors was tested on COS-7 cells. These CDs were modified with pyridylamino, alkylimidazole, methoxyethylamino or primary amine groups at 6-positions of the glucose units. Uncharged CDs, beta-CD, hydroxypropyl-beta-CD, and dimethyl-beta-CD were also tested, but these did not form stable complexes with the DNA and produced only a slight improvement in transfection level over DNA alone. The polycationic CDs neutralised DNA to form stable nanoparticulate complexes. The transfection efficiency of these CDs was dependent on the substituents present, with the most efficient having either an amino, pyridylamino or butylimidazole group at the 6-positions and unmodified 2- and 3-hydroxyls. One of the most effective vectors, heptakispyridylamino CD, produced a 4000-fold increase in transfection level over DNA alone. Levels were improved 10-fold by use of the endosomolytic agent, chloroquine. The transfection efficiency of the best of these systems in serum equals that of DOTAP in serum. Studies with (32)P-labelled plasmid DNA indicate that the polycationic CDs are exceptional promoters of DNA cellular-uptake, the most efficient surpassing DOTAP. Uptake is dependent on proteoglycan-mediated binding to cells. The data imply that intracellular trafficking but not cellular uptake, may be the rate-limiting step in the transfection process. These initial results indicate that CDs are useful templates for further modification to produce molecular constructs capable of enhanced gene delivery.
Cryan
Sally-Ann
SA
Department of Pharmaceutics and Pharmaceutical Technology, Trinity College, University of Dublin, Dublin 2, Ireland.
Holohan
Ann
A
Donohue
Ruth
R
Darcy
Raphael
R
O'Driscoll
Caitriona M
CM
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003505:Cyclodextrins / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism*; D000071228:Polyelectrolytes; D014162:Transfection / Q000379:methods*
D003505:Cyclodextrins; D011073:Polyamines; D000071228:Polyelectrolytes