Nanosheet transfection: effective transfer of naked DNA on silica glass

Abstract

Existing gene delivery technologies using nonviral vectors or physical stimulation are limited by cytotoxicity. Here, we reveal an easy and new method to fabricate silica upright nanosheets, which can be easily scaled up and used for gene delivery. We demonstrate that naked DNA can be transferred into difficult-to-transfect cells (for example, stem cells) by plating cells on silica glass with the upright nanosheets without using any vector. Gene entry is probably achieved through the integrin-FAK-Rho signaling axis, which can be activated by the cytoskeleton rearrangement on nanosheets in a limited time frame ('transfection window'). Transfecting naked GATA4-binding protein 4 plasmids into stem cells can upregulate the other two important cardiac marker genes myocyte enhancer factor 2C and T-box 5. The transfected mesenchymal stem cells express the cardiac marker proteins at 7 days after transfection, which confirms the success of the innovative gene delivery approach. The vector-free silica nanosheet-induced transfection is simple and effective but faster and safer than the conventional technologies.

Materials Nanoarchitectonics as Cell Regulators

“…Structural factor of the nanosheet, aspect ratio between height and thickness of the sheet walls, are significantly influential with transfection efficiency. Hsu and co‐workers also demonstrated a promoted transfection of naked plasmid DNA (GATA4) into human mesenchymal stem cells with more detailed gene expression analyses . The proposed method gives a higher gene transfection efficiency with a safe and naturally abundant silica as transfection substrates.…”

Two-Dimensional Nanosheet-Based Photonic Nanomedicine for Combined Gene and Photothermal Therapy

“…Huang at el. reported a way to achieve transfection via naked DNA on silica glass nanosheets (Huang et al, 2015). This method had unique advantages in a sense that no vector was needed, and naked DNA was able to be transferred into cells that were known to be difficult to transfect, which like stem cells.…”

Nanoarchitectonics for Hybrid and Related Materials for Bio‐Oriented Applications

“…Ji, Hsu, and co‐workers have achieved an easier fabrication of silica upright nanosheets using abundant silica glasses that could easily be scaled up . The as‐prepared silica nanosheets were used to demonstrate the transfer of naked DNA into difficult‐to‐transfect cells, such as stem cells.…”