Enhanced gene transfer into brain capillary endothelial cells using Antp-modified DNA-loaded nanoparticles.
Brain capillary endothelial cells (BCECs) have been considered as one of the primary targets for cerebral gene therapy. However, the cells, well-known for their poor function of endocytosis, are difficult to be transfected by general non-viral vectors. The aim of this study was to enhance the efficiency of transfection and expression in BCECs of DNA/polymer nanoparticles with the modification of membrane-penetrating peptide, Antennapedia peptide (Antp) polyethylenimine (PEI) and polyamidoamine (PAMAM) were chosen to prepare Antp-modified DNA-loaded nanoparticles with a complex coacervation technique. After a 20-min transfection, the efficiency, in terms of transfection and expression, of DNA/PEI NP or DNA/PAMAM NP was enhanced significantly with the modification of Antp. After a 3-h transfection of DNA/Antp/PEI NP, there was no difference in cellular uptake but an enhancement in gene expression, compared to DNA/PEI NP alone. However, both the transfection and expression efficiency of DNA/PAMAM NP were enhanced using Antp. These observations suggest that Antp can increase the membrane-penetrating ability of DNA-loaded nanoparticles, which can be employed as novel non-viral gene vectors.
Huang
Rong-Qin
RQ
Department of Pharmaceutics, School of Pharmacy, Fudan University, P.O. Box 232, Shanghai, 200032, China.
Pei
Yuan-Ying
YY
Jiang
Chen
C
D000818:Animals; D051943:Antennapedia Homeodomain Protein / Q000737:chemistry; D001921:Brain / Q000098:blood supply*; D002196:Capillaries / Q000166:cytology; D004247:DNA / Q000737:chemistry; D050091:Dendrimers; D004730:Endothelium, Vascular / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011073:Polyamines / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D051943:Antennapedia Homeodomain Protein; D050091:Dendrimers; C104700:PAMAM Starburst; D010455:Peptides; D011073:Polyamines; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA