Guanidinylated polymers enable safe and efficient dsRNA delivery in plant cells

Area of Science:

• Agricultural Science
• Polymer Chemistry
• Biotechnology

Background:

• Chemical pesticides pose environmental and health risks.
• RNA interference (RNAi) is a sustainable gene silencing alternative.
• dsRNA instability and poor cellular uptake limit practical RNAi application.

Purpose of the Study:

• To develop and evaluate polymethacrylate-based polymers for enhanced RNAi delivery in plants.
• To investigate the impact of polymer molar mass (MM) and degree of substitution (DS) on RNAi efficacy and safety.

Main Methods:

• Synthesis and characterization of twelve PAEMA and PGUMA polymers.
• Biophysical assays (nucleic acid condensation, DLS, z-potential) for polyplex evaluation.
• In vitro stability, cytotoxicity, cellular uptake, and gene silencing assays in plant systems.

Main Results:

• Polymers efficiently condensed nucleic acids into small, stable, positively charged polyplexes.
• Higher MM and DS polymers offered superior protection against degradation and reduced cytotoxicity.
• High DS polymers enhanced cellular uptake and siRNA delivery efficiency.
• RNAi delivery using these polymers resulted in effective transient gene silencing in plant cells.

Conclusions:

• Guanidinylated polymethacrylates are promising, biocompatible RNAi delivery vehicles for plants.
• Optimized polymer MM and DS balance stability, safety, and delivery efficiency.
• This approach presents a sustainable alternative to conventional chemical pesticides in agriculture.