Cross-linked polyethylenimine-hexametaphosphate nanoparticles to deliver nucleic acids therapeutics.
Branched polyethylenimine (PEI; 25 kDa) as a nonviral vector exhibits high transfection efficiency and is a potential candidate for efficient gene delivery. However, the cytotoxicity of PEI limits its application in vivo. PEI was ionically interacted with hexametaphosphate, a compact molecule with high anionic charge density, to obtain nanoparticles (PEI-HMP). Nanoparticles were assessed for their efficacy in protecting complexed DNA against nucleases. The intracellular trafficking of nanoparticles was monitored by confocal microscopy. The cytotoxicity and transfection efficiency of PEI-HMP nanoparticles were evaluated in vitro. In vitro transfection efficiency of PEI-HMP (7.7%) was approximately 1.3- to 6.4-folds higher than that of the commercial reagents GenePORTER 2, Fugene, and Superfect. Also, PEI-HMP (7.7%) delivered green fluorescent protein (GFP)-specific small interfering ribonucleic acid (siRNA) in culture cells leading to >80% suppression in GFP gene expression. PEI-HMP nanoparticles protected complexed DNA against DNase for at least 2 hours. A time-course uptake of PEI-HMP (7.7%) nanoparticles showed the internalization of nanoparticles inside the cell nucleus in 2 hours. Thus, PEI-HMP nanoparticles efficiently transfect cells with negligible cytotoxicity and show great promise as nonviral vectors for gene delivery.


BACKGROUND
Branched polyethylenimine (PEI) as a non-viral vector exhibits high transfection efficiency for gene delivery, but its cytotoxicity limits its applications. PEI hexametaphosphate nanoparticles (PEI-HMP) demonstrated a 1.3-6.4 folds higher transfection rate compared to commercial reagents. Overall, PEI-HMP nanoparticles efficiently transfect cells with negligible cytotoxicity and show great promise as non-viral vectors for gene delivery.
Patnaik
Soma
S
Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi, India.
Arif
Mohammad
M
Pathak
Atul
A
Kurupati
Raj
R
Singh
Yogendra
Y
Gupta
Kailash Chand
KC
D003432:Cross-Linking Reagents / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry*; D004339:Drug Compounding / Q000379:methods; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry*; D009696:Nucleic Acids / Q000008:administration & dosage* / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D003432:Cross-Linking Reagents; D004337:Drug Carriers; D009696:Nucleic Acids; D010710:Phosphates; C005035:hexametaphosphate; D011094:Polyethyleneimine