Graphene based gene transfection

Abstract

Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including biomedicine. In this work, for the first time we successfully use graphene as a non-toxic nano-vehicle for efficient gene transfection. Graphene oxide (GO) is bound with cationic polymers, polyethyleneimine (PEI) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI-10k polymer. The positively charged GO-PEI complexes are able to further bind with plasmid DNA (pDNA) for intracellular transfection of the enhanced green fluorescence protein (EGFP) gene in HeLa cells. While EGFP transfection with PEI-1.2k appears to be ineffective, high EGFP expression is observed using the corresponding GO-PEI-1.2k as the transfection agent. On the other hand, GO-PEI-10k shows similar EGFP transfection efficiency but lower toxicity compared with PEI-10k. Our results suggest graphene to be a novel gene delivery nano-vector with low cytotoxicity and high transfection efficiency, promising for future applications in non-viral based gene therapy.

Graphene‐Based MicroRNA Transfection Blocks Preosteoclast Fusion to Increase Bone Formation and Vascularization

“…Unbounded PEI was then removed by centrifugation and washing. Constructed GO‐PEI was stale in both saline (0.9% NaCl) cell medium without obvious agglomeration as reported 18. Since naked miRNAs are instable and easy to be catalyzed by enzymes, the best way for miRNA delivery is through a plasmid vector.…”

“…For GO‐PEI complexes preparation, PEI solution (1 mg mL −1 ) was slowly added into GO solution (0.1 mg mL −1 ) in 10 min. After ultrasonication for about 10 min, the mixture was stirred overnight, washed 3–5 times with deionized (DI) water by centrifugation, and then redispersed in DI water 18. GO‐PEI complexes were then mixed with miR‐7b GV159 plasmid in deionized water at the N/P ratios of 20.…”

“…Recently, it is reported that graphene oxide (GO) nanosheets, a precursor of graphene, can be used to deliver genes efficiently when ionically bonded to cationic polymers 17. We therefore used negatively charged GO to bind cationic polyethylenimine (PEI) polymers by electrostatic interactions, forming stable GO‐PEI complexes 18. PEI is known as a suitable material for gene transfer because its strongly binding to DNA, effectively uptaken by cells, demonstrating proton sponge effects, and helping delivered nucleic acids escape from the lysosomal pathways after cell internalization 19.…”

Graphene and graphene oxide as nanomaterials for medicine and biology application

“…Modified GO is used for gene delivery purposes. Polyethylenimine (PEI) modifies the surface of GO sheets and thus makes it ready for cellular gene delivery through covalent conjugation and electrostatic interaction [58] for plasmid DNA (pDNA) stacking. GO is attached covalently to the linear chain [35] and branched chain [59].…”

Environmental applications using graphene composites: water remediation and gas adsorption

“…These studies have shown the use of graphene in microbial fuel cells, 249 graphene as a scaffold for bacterial growth, 228 as well as the use of bacteria as a graphene oxide reducing agent. 250,251 Various in vitro toxicity studies have been conducted to determine the toxicity of graphene towards cancer [234][235][236][237][238] and human cell lines. 229,239 The cytotoxicity towards the cancer cell lines A549 was shown to depend largely on the cell culture medium components.…”