Defoliation and plasmid delivery with layer-by-layer coated colloids.
The uptake of polyelectrolyte multilayer coated colloids into cells, subsequent defoliation and plasmid delivery was studied by means of confocal microscopy and flow cytometry. Silica particles coated layer-wise with protamine and dextran sulfate were given to HEK 293T cells. Optimum uptake was found with protamine as the top layer. The particle uptake likely follows an non-receptor-mediated endocytotic pathway. Defoliation of polyelectrolyte multilayer coated particles within cells was demonstrated by the release of incorporated plasmids as indicated by the expression of plasmid encoded proteins using the enhanced green fluorescence proteine (pEGFP-C1) plasmid and a red fluorescence protein (pDsRed1-N1) plasmid. This proves, together with the direct observation of fluorescent layer debris, the defoliation of coated particles and the release of layer components into the cytoplasm. Particle uptake and GFP expression.
Reibetanz
Uta
U
Institute of Medical Physics and Biophysics, University of Leipzig, Härtelstrasse 16-18, 04107 Leipzig, Germany.
Claus
Claudia
C
Typlt
Elke
E
Hofmann
Jörg
J
Donath
Edwin
E
D000818:Animals; D001693:Biological Transport, Active; D002460:Cell Line; D002522:Chlorocebus aethiops; D003102:Colloids; D016264:Dextran Sulfate; D004337:Drug Carriers / Q000737:chemistry; D004573:Electrolytes; D005456:Fluorescent Dyes; D006801:Humans; D018613:Microscopy, Confocal; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011108:Polymers; D011479:Protamines; D013499:Surface Properties; D014709:Vero Cells
D003102:Colloids; D004337:Drug Carriers; D004573:Electrolytes; D005456:Fluorescent Dyes; D011108:Polymers; D011479:Protamines; D016264:Dextran Sulfate