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High-efficiency DNA injection into a single human mesenchymal stem cell using a nanoneedle and atomic force
Sung-Woong Han1, Chikashi Nakamura, Noriko Kotobuki
1Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, Japan.
Nanomedicine : Nanotechnology, Biology, and Medicine
|May 27, 2008
Summary
A novel nanoneedle, guided by atomic force microscopy (AFM), offers a low-invasive method for efficient gene delivery directly into cell nuclei. This technique surpasses traditional methods, achieving over 70% gene transfer in human mesenchymal stem cells.
Area of Science:
- Biotechnology
- Cell Biology
- Nanotechnology
Background:
- Traditional gene delivery methods often face challenges with efficiency and cellular damage.
- Microinjection and lipofection are common techniques but have limitations in delivery rates and invasiveness.
Purpose of the Study:
- To develop and evaluate a novel, low-invasive gene delivery system using an atomic force microscopy (AFM) nanoneedle.
- To assess the efficiency and cell compatibility of nanoneedle-mediated gene delivery into human mesenchymal stem cells (MSCs) and human embryonic kidney cells (HEK293).
Main Methods:
- An etched AFM tip (nanoneedle, 200 nm diameter, 6 µm length) was utilized for direct nuclear insertion.
- A plasmid encoding the green fluorescent protein (GFP) was adsorbed onto a poly-L-lysine-modified nanoneedle surface.
- The nanoneedle was inserted into primary cultured single human MSCs using an AFM system.
Main Results:
- Nanoneedle insertion probabilities were higher in MSCs and HEK293 cells compared to microinjection capillaries.
- Gene delivery efficiency exceeding 70% was achieved in human MSCs, significantly outperforming lipofection (~50%) and microinjection (~10%).
- The nanoneedle method yielded the highest expression of GFP per delivered DNA due to direct nuclear access and minimal cell damage.
Conclusions:
- The AFM nanoneedle represents a highly efficient and low-invasive tool for gene delivery directly into the cell nucleus.
- This method shows significant promise for applications in stem cell research and genetic engineering.
- The nanoneedle technology offers a superior alternative to existing nonviral gene delivery techniques.
