Parallel Synthesis and Screening of Polymers for Nonviral Gene Delivery

Abstract

We describe the parallel synthesis and in vitro evaluation of a cationic polymer library for the discovery of nonviral gene delivery vectors. The library was synthesized based on the ring-opening polymerization reaction between epoxide groups of diglycidyl ethers and the amines of (poly)amines. Parallel screening of soluble library constituents led to the identification of lead polymers with high DNA-binding efficacies. Transfection efficacies of lead polymers were evaluated using PC3-PSMA human prostate cancer cells and murine osteoblasts in the absence and presence of serum. In vitro experiments resulted in the identification of a candidate polymer that demonstrated significantly higher transfection efficacies and lower cytotoxicities than poly(ethyleneimine) (pEI), the current standard for polymeric transfection agents. In addition, polymers that demonstrated moderately higher and comparable transfection efficacies with respect to pEI were also identified. Our results demonstrate that high-throughput synthesis and screening of polymers is a powerful approach for the identification of novel nonviral gene delivery agents.

Comparison of Transfection Efficiency of Nonviral Gene Transfer Reagents

“…Therefore, it is important to carefully characterize the most suitable transfection system for each cell type prior to functional studies in order to achieve maximal transgene expression. Transfection efficiencies for MC3T3-E1 preosteoblasts have been reported using cationic liposome [12], cationic polymer [13], cationic lipid, gelatin, and PEI [14], nanostructured calcium phosphates [15], and gold nanoparticles [16]. In the present study, the highest transfection efficiency in MC3T3-E1 cells using luciferase gene in the presence and absence of serum was seen with FuGENE HD (lipids and other components).…”

Kinome-level screening identifies inhibition of polo-like kinase-1 (PLK1) as a target for enhancing non-viral transgene expression

“…The 1,4C-1,4bis polymer was synthesized by mixing the monomers 1,4-cyclohexane dimethanoldiglycidyl ether (1,4C, Sigma) and 1,4-bis(3-aminopropyl)-piperazine (1,4Bis, Sigma) at a 1:1 molar ratio as described previously [33]. 25 kDa branched polyethyleneimine (PEI) was purchased from Sigma.…”

“…PC3-PSMA cells were seeded in 96 well plates (15,000 cells/well) and incubated overnight (~18 h) at 37°C in serum-containing RPMI media (SCM). Polyplexes were prepared by mixing the polymer 1,4C-1,4Bis [33] with pGL3.0 plasmid DNA (Promega; luciferase expression plasmid with a SV40 promoter) at a 10:1 mass ratio and incubating the solution at room temperature for 20 min. SCM media was then removed from cells and replaced with SFM media.…”

Nanocombinatorial Chemistry in Nanomaterial Discovery and Nanomedicine

“…[46] 和免疫细胞 [47] 等均可产生细胞毒效应, 这些毒 性效应大大限制了它的应用范围. 研究表明, 通过表面 修饰功能化可以有效的调控纳米材料的生物活性 [26,48] . 我们在前期研究中, 证明了羧基化碳纳米管可以通过影 响 BMP 信号通路产生细胞周期阻滞, 影响细胞功能 [49] .…”