Plant genome editing is transforming agriculture by allowing scientists to make targeted genetic changes that improve crop yields, stress resistance, and nutritional value.1
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However, traditional gene delivery methods, such as Agrobacterium-mediated transformation and gene guns, are often slow, inefficient, and can damage plant tissues. These highlight the need for more precise and adaptable systems to deliver gene-editing tools like CRISPR/Cas9, RNA, and DNA.1,2
Nanotechnology offers a promising alternative. Nanoparticle-based delivery can enhance efficiency, bypass species-specific barriers, and work across a wider range of crops.
But how exactly does nanotechnology enable more effective genome editing, and what are the practical implications?
How Nanotechnology is Used in Genome Editing
Several types of nanoparticles have been used to deliver gene-editing tools.
Gold nanoparticles (AuNPs) are widely used due to their biocompatibility and ease of functionalization. AuNPs can bind to nucleic acids, protect them from degradation, and facilitate their delivery into plant cells. They have been successfully used for small interfering RNA (siRNA) delivery, achieving high gene knockdown efficiency in various plant species.3,4
Carbon nanotubes (CNTs)—both single-walled and multi-walled—are another promising nanocarrier. They are small, strong, and can penetrate plant cell walls.
Functionalized CNTs have been used to deliver DNA plasmids into plant cells, enabling transient gene expression. They can also electrostatically adsorb small RNA molecules, providing stability and enhancing cellular uptake while reducing off-target effects.5
Lipid-based nanoparticles (LNPs) are particularly effective for CRISPR/Cas9 delivery due to their high payload capacity, self-assembly properties, and ability to encapsulate ribonucleoprotein (RNPs).
Liposomes, a type of LNP, can be engineered with cationic lipids to facilitate the adsorption of negatively charged genetic material, such as sgRNA and Cas9 mRNA. This approach enhances genome-editing efficiency while minimizing toxicity and immune responses in plant cells.6
Mechanisms of Nanoparticle Delivery
Unlike traditional methods that rely on mechanical force or biological vectors, nanoparticles can enter plant cells through passive diffusion, endocytosis, or via plasmodesmata (tiny channels between plant cells). These routes allow nanoparticles to bypass structural barriers like the cell wall, reducing tissue damage and improving transformation efficiency.4
One key advantage of nanoparticle delivery is that it avoids the species-specific limitations of Agrobacterium-mediated transformation, which works well in dicots but poorly in many monocots like wheat and maize. Nanoparticles don’t rely on host-pathogen interactions and can be engineered to enter diverse plant species without the need for specific receptors.4
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Applications of Nanotechnology in Genome Editing
Case Study 1: Gold Nanoparticles for RNA-Based Gene Silencing
AuNPs have been widely investigated for their ability to deliver nucleic acids into plant cells. In a recent study by Lei et al., poly-disperse spherical AuNPs were used to transport siRNA into Arabidopsis thaliana protoplasts, achieving an 80 % gene knockdown efficiency.3
The siRNA-loaded AuNPs embedded in the plant cell wall successfully silenced target genes without requiring transgene integration. This method presents a promising alternative to traditional gene-silencing approaches, reducing off-target effects and improving precision in plant genome modification.3
Case Study 2: Carbon Nanotubes for DNA Delivery in Crops
CNTs are effective carriers for delivering genetic material to plants. In a study by Demirer et al., CNTs covalently grafted with polyethylenimine (PEI) were used to deliver a 4.2-kb DNA plasmid into Nicotiana benthamiana, arugula, wheat, and cotton.7
This allowed for the transient expression of green fluorescent protein (GFP), confirming successful gene delivery. Compared to traditional transformation techniques, CNT-mediated delivery bypasses the need for external mechanical force, enabling species-independent, high-efficiency DNA transfer.7
Case Study 3: Lipid-based Nanoparticles for CRISPR/Cas9 Delivery
In a study by Liu et al., Lipofectamine 3000—a commercial liposome transfection agent—was used for DNA-free delivery of Cas9 RNPs, achieving a 6 % editing efficiency. This was twice as effective as macro-gold biolistic methods.8
The ability of LNPs to encapsulate and protect gene-editing tools while ensuring targeted delivery enhances their potential for plant genetic engineering. This system overcomes challenges associated with traditional transformation techniques, offering a promising platform for developing transgene-free genome-edited crops.8
CRISPR Explained
Case Study 4: Silica Nanoparticles in Maize Genome Editing
Ortigosa et al. investigated mesoporous silica nanoparticles (MSNs) for DNA delivery in maize. Using biolistic methods, they delivered plasmids with MSNs sized between 100 and 200 nm. Uncapped MSNs proved ineffective, but capping them with gold nanoparticles significantly improved uptake.
The method achieved up to a 20 % increase in delivery efficiency and successfully produced herbicide-resistant maize, demonstrating the potential of MSNs in crop genome editing.9
Challenges and Ethical Considerations
The use of nanotechnology in genome editing faces significant regulatory uncertainty, as there are no clear guidelines for nano-based genetic modifications in agriculture.
Different countries adopt varying approaches. Some regulate based on the genetic state of the final product; others impose restrictions based on the modification process itself. The absence of standardized regulations creates ambiguity for researchers and industries seeking to commercialize nano-enabled gene-edited crops.10
Public skepticism toward genetically modified organisms (GMOs) extends to nano-enabled genome editing. While traditional GMOs involve inserting foreign DNA into a plant’s genome, nano-based gene editing primarily delivers transient modifications without integrating foreign genes.
This distinction could help avoid strict GMO regulations in some countries, but public concerns about food safety and environmental impact persist. Clear communication of the benefits and safety of nanoparticle-mediated gene editing is crucial for public acceptance.10,11
The long-term effects of nanoparticles in agriculture remain largely unexplored. Questions about nanoparticle degradation, potential toxicity, and accumulation in the environment or food chain need to be addressed through rigorous safety assessments. Additionally, the potential for unintended genetic modifications raises concerns about ecological stability, requiring long-term field studies to assess the broader implications.11
While nanoparticle-based genome editing has shown promising results in laboratories, commercial adoption faces challenges. Scalable production of functionalized nanoparticles, stability concerns, and delivery efficiency remain technical barriers. High costs may also limit accessibility, especially in developing countries.11
Bridging the gap to large-scale use will require further validation, regulatory approvals, and collaboration between researchers, policymakers, and industry stakeholders.
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References and Further Readings
1. Sharma, P.; Lew, TTS. (2022). Principles of Nanoparticle Design for Genome Editing in Plants. Frontiers in Genome Editing. https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2022.846624/full
2. Wu, K.; Xu, C.; Li, T.; Ma, H.; Gong, J.; Li, X.; Sun, X.; Hu, X. (2023). Application of Nanotechnology in Plant Genetic Engineering. International Journal of Molecular Sciences. https://pmc.ncbi.nlm.nih.gov/articles/PMC10573821/
3. Lei, W.-X.; An, Z.-S.; Zhang, B.-H.; Wu, Q.; Gong, W.-J.; Li, J.-M.; Chen, W.-L. (2020). Construction of Gold-Sirna Npr1 Nanoparticles for Effective and Quick Silencing of Npr1 in Arabidopsis Thaliana. Rsc Advances. https://pubs.rsc.org/en/content/articlelanding/2020/ra/d0ra02156c
4. Squire, H. J.; Tomatz, S.; Voke, E.; González-Grandío, E.; Landry, M. (2023). The Emerging Role of Nanotechnology in Plant Genetic Engineering. Nature Reviews Bioengineering. https://landrylab.com/wp-content/uploads/2023/02/aa8b1c60-b490-4629-acc2-59bd8d92b783.pdf
5. Ali, Z.; Serag, M. F.; Demirer, G. S.; Torre, B.; Di Fabrizio, E.; Landry, M. P.; Habuchi, S.; Mahfouz, M. (2022). DNA–Carbon Nanotube Binding Mode Determines the Efficiency of Carbon Nanotube-Mediated DNA Delivery to Intact Plants. ACS Applied Nano Materials. https://pubs.acs.org/doi/10.1021/acsanm.1c03482
6. Mahmoud, L. M.; Kaur, P.; Stanton, D.; Grosser, J. W.; Dutt, M. (2022). A Cationic Lipid Mediated Crispr/Cas9 Technique for the Production of Stable Genome Edited Citrus Plants. Plant methods. https://plantmethods.biomedcentral.com/articles/10.1186/s13007-022-00870-6
7. Demirer, G. S.; Zhang, H.; Matos, J. L.; Goh, N. S.; Cunningham, F. J.; Sung, Y.; Chang, R.; Aditham, A. J.; Chio, L.; Cho, M.-J. (2019). High Aspect Ratio Nanomaterials Enable Delivery of Functional Genetic Material without DNA Integration in Mature Plants. Nature nanotechnology. https://pubmed.ncbi.nlm.nih.gov/30804481/
8. Liu, W.; Rudis, M. R.; Cheplick, M. H.; Millwood, R. J.; Yang, J.-P.; Ondzighi-Assoume, C. A.; Montgomery, G. A.; Burris, K. P.; Mazarei, M.; Chesnut, J. D. (2020). Lipofection-Mediated Genome Editing Using DNA-Free Delivery of the Cas9/Grna Ribonucleoprotein into Plant Cells. Plant cell reports. https://pubmed.ncbi.nlm.nih.gov/31728703/
9. Martin-Ortigosa, S.; Peterson, D. J.; Valenstein, J. S.; Lin, V. S.-Y.; Trewyn, B. G.; Lyznik, L. A.; Wang, K. (2014). Mesoporous Silica Nanoparticle-Mediated Intracellular Cre Protein Delivery for Maize Genome Editing Via Loxp Site Excision. Plant physiology. https://pmc.ncbi.nlm.nih.gov/articles/PMC3912087/
10. Vats, S.; Kumawat, S.; Brar, J.; Kaur, S.; Yadav, K.; Magar, S. G.; Jadhav, P. V.; Salvi, P.; Sonah, H.; Sharma, S. (2022). Opportunity and Challenges for Nanotechnology Application for Genome Editing in Plants. Plant Nano Biology. https://www.sciencedirect.com/science/article/pii/S2773111122000018
11. Khan, S. H. (2019). Genome-Editing Technologies: Concept, Pros, and Cons of Various Genome-Editing Techniques and Bioethical Concerns for Clinical Application. Molecular therapy Nucleic acids. https://pubmed.ncbi.nlm.nih.gov/30965277/
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