Saponin and fluorine-modified polycation as a versatile gene delivery system

Abstract

Despite the development of many novel carriers for the delivery of various types of genetic material, the lack of a delivery system with high efficiency and low cytotoxicity is a major bottleneck. Herein, low molecular weight polyethylenimine (PEI1.8k) was functionalized with saponin residues using phenylboronic acid (PBA) as an ATP-responsive cross-linker, and a fluorinated side chain to construct PEI-PBA-SAP-F polycation as a highly efficient delivery vector. This vehicle could transfect small plasmid DNA ( 3 kb) with outstanding efficiency into various cells, including HEK293T, NIH3T3, A549, PC12, MCF7 and HT-29, as well as robust transfection of a large plasmid ( 9 kb) into HEK293T cells. The carrier could maintain transfection efficacy even at high concentration of serum and low doses of plasmid. The use of green fluorescent protein (GFP) knock-out analysis demonstrated transfection of different types of CRISPR/Cas9 complexes (Cas9/sgRNA ribonucleoproteins RNP, plasmid encoding Cas9 plus sgRNA targeting GFP, Cas9 expression plasmid plus in vitro-prepared sgRNA). The transfection of a large plasmid into local mouse brain tissue was also shown. In summary, we report an effective PEI-PBA-SAP-F gene carrier with the appropriate lipophilic/cationic balance for biomedical applications.

Combination of anti-miR19a-3p polyplex plus doxorubicin for breast cancer in 2D culture and apoptosis assay in 3D spheroids in a microwell device

“…We used PPSF polymeric nanoparticles for the delivery of anti-miR19a-3p into cell lines. The smart PPSF NPs were prepared as described in our previous study [38]. Briefly, NHS, EDC and PBA were dissolved in dry methanol, then mixed slightly at room temperature (RT) for 24 h (h).…”

“…The culture media of each plate was replaced with fresh medium without serum and Pen/Strep. As described in our previous study [38], we used PPSF polymers in 20X weight ratios, as follows: scramble or anti-miR19a-3p (1 µg) in 25 µl hepes buffer and PPSF polymers (2 µl of 1 mg/100 µl stock solution) in 25 µl hepes buffer were prepared, then PPSF polymer solution was added to scramble or anti-miR19a-3p solution, incubated for 30 min at RT, and added to each plate. After 4 h incubation time had elapsed, the medium was replaced with fresh medium with 10% FBS and 1% Pen/Sterp.…”

“…Our previous report described smart PEI-PBA-SAP-F 15 (PPSF) polymeric nanoparticles, and demonstrated that these PPSF polymers could efficiently deliver genetic materials into a variety of cultured cell lines [38]. The first reason for this excellent transfection efficiency was attributed to the presence of the saponin moiety, which can improve transfection by interacting with cholesterol in the cell membrane, and destabilize the lysosomal membrane leading to endosomal escape.…”

“…The second reason was attributed to the presence of the fluorine substitution. Fluorine substitution can improve the transport of polymeric nanoparticle cargos in three ways: (1) increasing cell uptake and endosomal escape; (2) reducing the surface charge; (3) improving the plasmid release rate [38].…”

Gold Nanoparticles as Drug Carriers: The Role of Silica and PEG as Surface Coatings in Optimizing Drug Loading

“…There are many different types of drug delivery systems, including micro- [ 2 ] and nanoparticles [ 3 ], nanoplatforms [ 4 , 5 , 6 ], devices [ 7 , 8 , 9 , 10 ], and theragnostic platforms [ 11 , 12 , 13 ]. These systems can be used to deliver a wide range of drugs, including chemotherapeutics [ 14 , 15 , 16 ], gene therapies [ 17 , 18 , 19 , 20 ], and other biologics [ 21 , 22 , 23 , 24 , 25 , 26 ]. Optimizing drug loading in drug delivery systems is a critical aspect of their design and performance.…”