Disulfide‐bond‐containing agamatine‐cystaminebisacrylamide polymer demonstrates better transfection efficiency and lower cytotoxicity than polyethylenimine in NIH/3T3 cells

Abstract

Previously, we synthesized a non-viral vector containing disulfide bond by polymerization of agamatine (AGM) and N,N'-cystaminebisacrylamide (CBA). In this study, we investigated the transfection efficiency of disulfide bond (SS) containing AGM-CBA polymer in gene delivery into NIH/3T3 cells, and examined the factors affecting its transfection efficiency by comparing with polyethylenimine (PEI). In addition, experiments were carried out to determine the mechanisms of cell entry pathways and intracellular behavior of AGM-CBA/pDNA polyplexes. The transfection efficiency of AGM-CBA/pDNA with different weight ratios and different amounts of pDNA was measured and the pathways mediated transfection processes were studied by using various endocytosis inhibitors. To determine the intracellular behavior of AGM-CBA/pDNA polyplexes, the transfection efficiencies of AGM-CBA/pDNA and PEI/pDNA polyplexes with different combination structures were determined by using reporter gene and fake plasmid DNA. The transfection efficiency of AGM-CBA/pDNA polyplexes was correlated with its weight ratio of AGM-CBA and pDNA, and the amount of pDNA. Both AGM-CBA/pDNA and PEI/pDNA polyplexes enter into cell by clathrin- and caveolae-mediated endocytic pathways. However, AGM-CBA/pDNA showed different intracellular behavior in NIH/3T3 cells compared to PEI/pDNA polyplexes. It was hypothesized that disulfide bond in AGM-CBA could be an important factor contributing to its intracellular behavior and better transfection efficiency. Overall, AGM-CBA demonstrated better transfection efficiency and lower cytotoxicity than PEI in NIH/3T3 cells as a gene delivery vector.

Inhibition of Tumor Growth via Systemic siRNA Delivery Using Reducible Bile Acid-Conjugated Polyethylenimine

“…TP(3)-DA-PEI/thiol-siRNA demonstrated superior performance of cellular delivery of siRNA among the formulations (Figure 6). The enhanced cellular delivery efficiency of polyplexes with tight and compact structures using disulfide linkages can also be found in previous observations [28,29]. The limited stability of polyplex because of the short and rigid structure of siRNA could be improved by the formation of the disulfide linkages between thiol-siRNA and TP(3)-DA-PEI.…”

Nuclear delivery of plasmid DNA determines the efficiency of gene expression

“…** P < 0.01 compared with each other. Data obtained from previous report (Yang et al, ). (H) Ratios of transfection expression of AGM‐CBA/pLUC and PEI/pLUC polyplexes to the pLUC amount delivered in nuclei at 8 h.…”

“…To build on previous work (Yu et al, ; Yang et al, ), we used a quantitative polymerase chain reaction (qPCR) method to determine pLUC distribution in whole cells and nuclei at different time points after transfection studies of AGM‐CBA/pLUC and PEI/pLUC in NIH/3T3 cells. The intracellular trafficking properties of AGM‐CBA/pLUC and PEI/pLUC were compared by measuring pLUC amount in whole cells at 4 h and nuclei at 8 h, respectively.…”

Polyamidoamines: Versatile Bioactive Polymers with Potential for Biotechnological Applications

“…Linear bioreducibles PAA with different steric hindrance, hence with diversified chemical stability, near the disulfide were synthesized (Scheme 10a) [119]. Bioreducible PAAs with increased cell penetration ability were obtained by introducing guanidine pendants either by homopolymerizing N,N'-cystaminebisacrylamide with 4-aminebutylguanidine [120] or by copolymerizing N,N'-cystaminebisacrylamide with 4-aminebutanol/4-aminebutylguanidine [121] or hystidine/4-aminebutylguanidine mixtures [122].…”