Abstract
We recently developed a polyethylenimine (PEI) and polyethylene glycol (PEG) dual-functionalized reduced graphene oxide (GO) (PEG−nrGO−PEI, RGPP) for high-efficient gene delivery in HepG2 and Hela cell lines. To evaluate the feasibility and applicability of RGPP as a gene delivery carrier, we here assessed the transfection efficiency of RGPP on gene plasmids and siRNA in 11 different cell lines. Commercial polyalkyleneimine cation transfection reagent (TR) was used as comparison. In HepG2 cells, RGPP exhibited much stronger delivery ability for siRNA and large size plasmids than TR. For green fluorescent protein (GFP) plasmid, RGPP showed about 47.1% of transfection efficiency in primary rabbit articular chondrocytes, and about 27% of transfection efficiency in both SH-SY5Y and A549 cell lines. RGPP exhibited about 37.2% of GFP plasmid transfection efficiency in EMT6 cells and about 26.0% of GFP plasmid transfection efficiency in LO2 cells, but induced about 33% of cytotoxicity in both cell lines. In 4T1 and H9C2 cell lines, RGPP had less than 10% of GFP plasmid transfection efficiency. Collectively, RGPP is a potential nano-carrier for high-efficiency gene delivery, and needs to be further optimized for different cell lines.
Highlights
Gene delivery provides a powerful tool for exploring gene function, generating transgenic organisms and treating gene-related diseases [1,2,3]
Complexes of RGPP/siRNA or transfection reagent (TR)/siRNA were incubated with cells for 4 h in 500 μl serum-free media, and siRNA transfection efficiency was determined by fluorescence microscope imaging
After exposure of cells to RGPP or TR for 48 h, Cell Counting Kit-8 (CCK-8) assay showed that both RGPP and TR exhibited dose-dependent cytotoxicity, and RGPP at N/P ratio of 60 and 0.4% (TR volume/medium volume) of TR had similar cytotoxicity
Summary
Gene delivery provides a powerful tool for exploring gene function, generating transgenic organisms and treating gene-related diseases [1,2,3]. CRISPR/Cas, a versatile genome editing technology, can target nearly any DNA sequence, and the high efficiency of genome editing with Cas makes it possible to alter many targets in parallel, thereby enabling unbiased genome-wide functional screens to identify genes that play an important role in a phenotype of interest [15,16,17]. CRISPR/Cas can generate cellular transgenic models for studying human polygenic diseases, generating transgenic animal models and repressing or activating gene transcription [15,18,19,20,21,22,23]. With high potential to treat diseases at genetic roots, gene therapy has long fascinated scientists and clinicians. In the past two decades, a series of phase I/II gene-therapy clinical trials have been reported to have significant efficacy and safety for the treatment of various severe inherited diseases of the immune, blood and nervous systems [24,25,26,27,28]
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