Sterically stabilized polyplex: ligand-mediated activity.
Synthetic vectors have been considered as a safer and more versatile alternative to viral-based gene delivery systems. A variety of very simple synthetic vector systems, e.g., cationic lipid- and polymer-complexed plasmid DNA have activity in vivo but it appears to be mediated by non-specific electrostatic interactions limiting targeting. In order to avoid these problems, we designed a sterically stabilized layered colloidal system. The steric polymer coating reduces non-specific interactions. We have synthesized a PEG conjugate of PEI that complexes DNA to form small, stable colloids with a steric polymer coat on their surface. The polymer enhances colloidal stability and reduces non-specific binding and toxicity. It also renders the complex inactive presumably due to reduced binding. Ligands are then appended to the distal end of the steric polymer to restore cell binding and expression at target cells. We prepared conjugates with RGD peptide ligands appended to the distal end of the steric polymer. The resulting conjugates also form complexes but with ligands exposed on their surface restoring binding and activity. Labeled oligonucleotides and DNA were used to measure intracellular distribution. Oligonucleotides are found localized in the nucleus, whereas the labeled plasmid DNA remained in the cytoplasm. Import of plasmid DNA into the nucleus appears to be very inefficient yet sufficient for expression.
Woodle
M C
MC
Intradigm Corporation, 8205 Beech Tree Road, Bethesda, MD 20817, USA. mwoodle@intradigm.com
Scaria
P
P
Ganesh
S
S
Subramanian
K
K
Titmas
R
R
Cheng
C
C
Yang
J
J
Pan
Y
Y
Weng
K
K
Gu
C
C
Torkelson
S
S
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D005822:Genetic Vectors / Q000737:chemistry* / Q000494:pharmacology; D008024:Ligands; D008156:Luciferases / Q000235:genetics; D010316:Particle Size; D010957:Plasmids; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D011108:Polymers
D008024:Ligands; D011108:Polymers; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D008156:Luciferases