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Polymer-Coated Hydroxyapatite Nanocarrier for Double-Stranded RNA Delivery
Zeinah Elhaj Baddar1, Dhandapani Gurusamy1, Jérôme Laisney2
1Department of Entomology, University of Kentucky, Lexington, Kentucky 40546, United States.
Journal of Agricultural and Food Chemistry
|June 4, 2020
Summary
Polymer-functionalized hydroxyapatite nanoparticles effectively deliver double-stranded RNA (dsRNA) for insect pest control, significantly improving gene silencing compared to naked dsRNA in cellular studies.
Area of Science:
- Nanotechnology in Pest Management
- Molecular Biology and Genetics
- Biochemistry and Materials Science
Background:
- Conventional insecticides face challenges like insect resistance and environmental harm.
- RNA interference (RNAi) shows promise for pest control, but naked double-stranded RNA (dsRNA) has limited efficacy.
- Nanocarriers offer a strategy to enhance dsRNA delivery and effectiveness.
Purpose of the Study:
- To develop and evaluate polymer-functionalized hydroxyapatite (HA) nanoparticles as nanocarriers for dsRNA.
- To assess the efficacy of these nanocarriers in achieving gene silencing in insect cells.
- To investigate the cellular uptake and delivery mechanism of dsRNA via nanocarriers.
Main Methods:
- Synthesis of poly(acrylic acid) (PAA)-coated HA nanoparticles (PAA-HA NPs).
- Coating of PAA-HA NPs with poly-l-arginine (PLR10) for dsRNA binding via electrostatic assembly.
- In vitro gene silencing assays using transgenic SF9 cells and firefly luciferase reporter gene.
- Confocal microscopy to study cellular localization of dsRNA.
Main Results:
- Binding of dsRNA to PLR10-PAA-HA NPs increased with higher nanoparticle-to-dsRNA ratios.
- Significant gene silencing (35% decrease) was achieved with PLR10-PAA-HA NPs at a 5:1 ratio, while naked dsRNA was ineffective.
- A concentration-response relationship was observed for gene knockdown, with cytotoxicity at high concentrations.
- Confocal microscopy revealed dsRNA from nanocarriers bypassed endosomal entrapment, unlike naked dsRNA.
Conclusions:
- Polymer-functionalized HA nanocarriers effectively deliver dsRNA for gene knockdown in insect cells.
- This nanocarrier system overcomes the limitations of naked dsRNA, offering a promising approach for RNAi-based pest control.
- Further research is warranted to optimize delivery and assess in vivo efficacy and safety.
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