Preparation and evaluation of a new class of gene transfer reagents: poly(-alkylaminosiloxanes).
We report the evaluation of poly(-alkylaminosiloxane) as a novel class of polycationic DNA carriers. Controlled hydrolysis of mono- and di-aminoalkylmethyldimethoxysilane provided a wide range of defined oligomeric mixtures. Basic hydrolysis conditions yielded mixtures composed mainly of cyclic and long linear oligomers, while under acidic conditions mainly short-linear oligomers were derived. They all efficiently interacted with plasmid DNA as revealed by electron microscopy and DNA retardation assays. However, only diamine-based oligomers prepared under basic conditions were able to mediate substantial levels of DNA transfection in human HeLa cells. SiDA1b, prepared by basic hydrolysis of 3-(2-aminoethylamino)propyl-methyl-dimethoxysilane, was found to be at least as efficient as the frequently used cationic transfection agents DOTAP and polyethylenimine (PEI). The transfection activity was sensitive to bafilomycin A1, suggesting a mechanism that depends on proton capture during the acidification process associated with endocytosis.
Kichler
Antoine
A
Genethon-CNRS UMR 8115, 1 Rue de l'Internationale F-91002 Evry, France. akich@genethon.fr
Sabourault
Nicolas
N
Décor
Rachel
R
Leborgne
Christian
C
Schmutz
Marc
M
Valleix
Alain
A
Danos
Olivier
O
Wagner
Alain
A
Mioskowski
Charles
C
D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D011817:Rabbits; D012833:Siloxanes / Q000008:administration & dosage* / Q000138:chemical synthesis*
D012833:Siloxanes; D004247:DNA