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DNA Delivery by Virus-Like Nanocarriers in Plant Cells
Md Reyazul Islam1, Marina Youngblood1, Hye-In Kim1
1Department of Botany and Plant Sciences, University of California, Riverside, California 92507, United States.
Nano Letters
|June 18, 2024
Summary
Tobacco mild green mosaic virus (TMGMV)-like nanocarriers, coated with poly(allylamine) hydrochloride (PAH), efficiently deliver DNA to plant cells. These biocompatible nanocarriers facilitate gene expression in plants, advancing genetic engineering applications.
Area of Science:
- Plant biotechnology
- Nanotechnology
- Molecular biology
Background:
- Developing efficient and safe gene delivery systems is crucial for plant genetic engineering.
- Virus-like particles offer a promising platform for nanocarrier development due to their structural properties.
- Biocompatible materials are essential for minimizing off-target effects in plant cells.
Purpose of the Study:
- To design and characterize TMGMV-PAH nanocarriers for plant gene delivery.
- To evaluate the efficiency of TMGMV-PAH in delivering DNA to plant cells and facilitating gene expression.
- To assess the safety and biocompatibility of the nanocarriers for potential in vivo applications.
Main Methods:
- High aspect ratio Tobacco mild green mosaic virus (TMGMV) were coated with poly(allylamine) hydrochloride (PAH).
- Nanocarrier charge and DNA loading capacity were analyzed.
- Gene delivery efficiency was tested in Arabidopsis protoplasts using plasmid DNA encoding GFP.
- Inactivated TMGMV-PAH (iTMGMV-PAH) were used for in vivo studies, with gene expression analyzed via confocal microscopy and RT-qPCR.
Main Results:
- TMGMV-PAH nanocarriers exhibited a high surface charge (56.20 ± 4.7 mV) and efficiently loaded DNA.
- TMGMV-PAH facilitated DNA uptake via energy-independent mechanisms in Arabidopsis protoplasts.
- Successful delivery of plasmid DNA encoding GFP to the protoplast nucleus resulted in significant GFP expression (70% viability).
- Inactivated iTMGMV-PAH demonstrated effective in vivo gene delivery and expression of GFP in plants.
Conclusions:
- TMGMV-PAH nanocarriers are effective, biocompatible tools for plant gene delivery.
- The energy-independent uptake mechanism highlights the nanocarriers' potential for broad application.
- UV inactivation of nanocarriers prevents systemic infection, ensuring safety for in vivo use.
- Virus-like nanocarrier-mediated gene delivery represents a facile approach for advancing plant genetic engineering.
