WRAP-based nanoparticles for siRNA delivery in zebrafish embryos by simple bath immersion

Area of Science:

• Biotechnology
• Molecular Biology
• Zebrafish Research

Background:

• RNA interference (RNAi) is a powerful gene silencing tool in biomedical research.
• Effective delivery of small interfering RNA (siRNA) is critical for RNAi success, especially in vivo.
• Cell-penetrating peptides, like WRAP, are promising candidates for siRNA delivery systems.

Purpose of the Study:

• To optimize WRAP-based nanoparticles for efficient siRNA delivery and gene silencing in zebrafish embryos.
• To determine the ideal formulation of WRAP1 nanoparticles compatible with zebrafish embryo care conditions.
• To demonstrate the efficacy of PEGylated WRAP1 nanoparticles for in vivo gene silencing via skin transfection.

Main Methods:

• Formulation of WRAP1 nanoparticles with varying concentrations of polyethylene glycol (PEG).
• Characterization of nanoparticle size and uniformity (polydispersity index).
• Transfection of dechorionated zebrafish embryos expressing GFP using siRNA-loaded PEG-WRAP1 nanoparticles via a soaking method.

Main Results:

• Addition of 20% PEG to WRAP1 nanoparticles resulted in optimal size (~100 nm) and uniformity (PdI ≤0.3).
• siRNA-loaded 20% PEG-WRAP1 nanoparticles were efficiently internalized by zebrafish embryos.
• Dose-dependent and efficient GFP gene silencing was observed in zebrafish embryos.

Conclusions:

• PEGylated WRAP1 nanoparticles represent a promising delivery system for siRNA in zebrafish embryos.
• This optimized formulation facilitates effective in vivo gene silencing through simple skin transfection.
• Further research in zebrafish models can explore the therapeutic potential of these nanoparticles in pathophysiological contexts.