Gene delivery to differentiated neurotypic cells with RGD and HIV Tat peptide functionalized polymeric nanoparticles.
A number of neurodegenerative disorders may potentially be treated by the delivery of therapeutic genes to neurons. Nonviral gene delivery systems, however, typically provide low transfection efficiency in post-mitotic differentiated neurons. To uncover mechanistic reasons for this observation, we compared gene transfer to undifferentiated and differentiated SH-SY5Y cells using polyethylenimine (PEI)/DNA nanocomplexes. Differentiated cells exhibited substantially lower uptake of gene vectors. To overcome this bottleneck, RGD or HIV-1 Tat peptides were attached to PEI/DNA nanocomplexes via poly(ethylene glycol) (PEG) spacer molecules. Both RGD and Tat improved the cellular uptake of gene vectors and enhanced gene transfection efficiency of primary neurons up to 14-fold. RGD functionalization resulted in a statistically significant increase in vector escape from endosomes, suggesting it may improve gene delivery by more than one mechanism.
Suk
Jung Soo
JS
Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, MD 21218, USA.
Suh
Junghae
J
Choy
Kokleong
K
Lai
Samuel K
SK
Fu
Jie
J
Hanes
Justin
J
D001672:Biocompatible Materials; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D015696:Gene Products, tat; D018014:Gene Transfer Techniques; D006678:HIV / Q000235:genetics; D006801:Humans; D009474:Neurons / Q000166:cytology; D009842:Oligopeptides; D010446:Peptide Fragments / Q000235:genetics; D054322:tat Gene Products, Human Immunodeficiency Virus
D001672:Biocompatible Materials; D015696:Gene Products, tat; D009842:Oligopeptides; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; C047981:arginyl-glycyl-aspartic acid