Nucleic acid delivery with chitosan hydroxybenzotriazole.
The objective of this study was to investigate the transfection efficiency of chitosan hydroxybenzotriazole (CS-HOBT) for in vitro nucleic acid delivery. The results revealed that CS-HOBT was able to condense with DNA/small interfering double-stranded RNA molecules (siRNA). Illustrated by agarose gel electrophoresis, complete complexes of CS-HOBT/DNA were formed at a weight ratio of above 3, whereas those of CS-HOBT/siRNA were formed at a weight ratio of above 4 (CS molecular weights [MWs] 20 and 45 kDa) and above 2 (CS MWs 200 and 460 kDa). Gel electrophoresis results indicated that binding of CS-HOBT and DNA or siRNA depended on the MW and weight ratio. The particle sizes of CS-HOBT/nucleic acid complexes were in nanosize range. The highest transfection efficiency of CS-HOBT/DNA complex was found at a weight ratio of 2, with the lowest CS MW of 20 kDa. The CS-HOBT-mediated siRNA silencing of the enhanced green fluorescent protein gene occurred maximally with 60% efficiency. The CS-HOBT/siRNA complex with the lowest CS MW of 20 kDa at a weight ratio of 80 showed the strongest inhibition of gene expression. For cytotoxicity studies, over 80% the average cell viabilities of the complexes were observed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. This study suggests that CS-HOBT is straightforward to prepare, is safe, and exhibits significantly improved nucleic acid delivery potential in vitro.
Opanasopit
Praneet
P
Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, Thailand. praneet@su.ac.th
Techaarpornkul
Sunee
S
Rojanarata
Theerasak
T
Ngawhirunpat
Tanasait
T
Ruktanonchai
Uracha
U
D002460:Cell Line; D048271:Chitosan / Q000737:chemistry*; D004337:Drug Carriers; D006801:Humans; D009696:Nucleic Acids / Q000008:administration & dosage*; D010957:Plasmids; D034741:RNA, Small Interfering
D004337:Drug Carriers; D009696:Nucleic Acids; D034741:RNA, Small Interfering; D048271:Chitosan