J DRUG DELIV SCI TEC. 2022; 75, DOI:10.1016/j.jddst.2022.103669

Development of a non-viral gene vector for enhancing gene transfection efficiency

Li, Y; Yu, T; Han, LZ; Jin, LL; Jin, Y; Quan, JS

Abstract

Construction of a safe and high transfection efficiency non-viral gene delivery vector to improve the effectiveness of gene delivery is vital for cancer treatment. Herein, we used 1, 2-Dioleoyl-3-Trimethylammonium-Propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol to prepare cationic lipo-somes in combination with poly(aspartate-graft-PEI423) (PAE) and plasmid DNA to prepare Liposomes-poly (aspartate-graft-PEI423)/DNA (LP-PAE/DNA) to improve gene transfection efficiency. LP-PAE/DNA particle size was about 220 nm, the zeta potential was about 20 mV and the morphology was spherical. The LP-PAE/DNA lipopolyplexes showed higher transfection efficiency at a weight ratio of 10:1:1 in HeLa, A549 and SPC-A1 cell line. LP-PAE/DNA lipopolyplexes toxicity in different cell lines and the main endocytic pathways through which LP and LP-PAE mediate DNA into cells was examined. In addition, the results of hemolysis test, red blood cell morphology observation and blood routine experiments showed that LP-PAE/DNA would not cause systemic hematological toxicity. The histological analysis including hematoxylin-eosin (HE) staining and terminal deox-ynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay confirmed lipopolyplexes/li-posomes mediate the entry of therapeutic genes into the body not causing damage to normal organs and enhancing tumor tissue apoptosis relative to more traditional approaches. Therefore, it could be hoped the feasibility of LP-PAE as an efficient and promising platform for gene delivery.

Keywords: Lipopolyplexes; Cytotoxicity; Transfection efficiency; Endocytosis

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Transfection

Nanoparticles play a crucial role in the process of mRNA transfection, a cutting-edge technology that utilizes these particles as carriers to transport mRNA into cells for the purpose of achieving protein expression and gene editing. Through surface modification, nanoparticles can ensure both efficient stability and cell-specific delivery of mRNA. This transfection technique not only sidesteps the safety concerns associated with traditional viral vectors but also enhances delivery efficiency. The size and surface charge of nanoparticles can be manipulated to regulate their properties during cellular uptake and internal release. Moreover, the functionalization of nanoparticle surfaces enables targeted delivery, facilitating the precise delivery of mRNA to specific cells or tissues. mRNA transfection, facilitated by nanoparticles, introduces innovative approaches for gene therapy, vaccine development, and cell therapy, among other fields. This technology holds significant value for swiftly responding to infectious diseases, advancing cancer treatment, and advancing personalized medicine. It paves the way for novel avenues in biomedical research and treatment methods, marking a noteworthy contribution to the promotion of innovation in the realm of healthcare.

Product Name	Catalog	Unit Size	Price
Hela Transfection Reagent	WHM-OB12	40 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
Cos Transfection Reagent	WHM-OB13	50 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
BiochemFect Transfection Reagent	WHM-OB14	50 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
BiochemFect Gold Transfection Reagent	WHM-OB15	40 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
BiochemFect Stem Transfection Reagent	WHM-OB16	50 µL, 500 µL, 1 mL	INQUIRY
EcoFect Transfection Reagent	WHM-OB17	50 µL, 500 µL, 1 mL, 3x 1 mL	INQUIRY
lMagFec Transfection Reagent	WHM-OB21-1	200 µL, 500 µL, 1 mL, 3x 1 mL	INQUIRY
siRNA Transfection Selection Kit	WHM-OB2A	1 Kit	INQUIRY
Silent siRNA Transfection Reagent	WHM-OB22	50 µL, 500 µL, 1 mL, 3x 1 mL	INQUIRY
Silent Stem siRNA Transfection Reagent	WHM-OB23	50 µL, 500 µL, 1 mL	INQUIRY
mRNA Transfection Reagent	WHM-OB24	50 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
Stem mRNA Transfection Reagent	WHM-OB25	50 µL, 500 µL, 1 mL, 5x 1 mL	INQUIRY
mRNA/gRNA Transfection Reagent	WHM-OB26	50 µL, 500 µL	INQUIRY
Golden Tran-D	CDSNT42	0.1 mL, 1.5 mL, 1.0 mL, 0.5 mL, 2.0 mL, 5.0 mL, 3.0 mL	INQUIRY
Golden Tran-DR	CDSNT43	0.1 mL, 1.5 mL, 1.0 mL, 0.5 mL, 2.0 mL, 5.0 mL, 3.0 mL	INQUIRY
Golden Tran-R	CDSNT45	0.1 mL, 1.5 mL, 1.0 mL, 0.5 mL, 2.0 mL, 5.0 mL, 3.0 mL	INQUIRY
Palmitate Phosphoramidite	CDGLR01	50 µmol, 100 µmol, 0.25 g	INQUIRY
Stearyl Phosphoramidite	CDGLR04	100 µmol, 0.25 g	INQUIRY
5'-Cholesteryl-TEG Phosphoramidite	CDGLR06	50 µmol, 100 µmol, 0.25 g	INQUIRY
Cholesteryl-TEG Phosphoramidite	CDGLR07	50 µmol, 100 µmol, 0.25 g	INQUIRY

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