Designing siRNA/chitosan-methacrylate complex nanolipogel for prolonged gene silencing effects

Area of Science:

• Biotechnology
• Nanomedicine
• Gene Therapy

Background:

• Sustained release of small interfering RNA (siRNA) is crucial for effective gene therapy but remains challenging.
• Nanoparticle-based delivery systems offer a promising strategy to improve siRNA therapeutic efficacy.

Purpose of the Study:

• To develop UV crosslinkable nanolipogels (NLGs) encapsulating siRNA/chitosan-methacrylate (CMA) complexes for sustained siRNA release.
• To investigate the cellular uptake mechanisms and transfection efficiency of the developed NLGs.

Main Methods:

• Encapsulation of siRNA/CMA complexes into liposomes to form NLGs.
• In vitro assessment of siRNA release kinetics over 28 days.
• Investigation of cellular entry pathways using endocytic inhibitors.
• Evaluation of transfection efficacy and gene silencing duration in cell cultures.

Main Results:

• NLGs demonstrated enhanced siRNA encapsulation efficiency.
• Sustained siRNA release was observed for up to 28 days in vitro.
• Positively charged NLGs entered cells via multiple endocytosis pathways, facilitating endosomal escape and cytoplasmic siRNA release.
• Effective transfection and prolonged gene silencing up to 14 days were achieved in cell cultures.

Conclusions:

• The developed siRNA-loaded nanolipogel platform provides sustained release and effective gene silencing.
• This NLG system shows potential for both fundamental research and clinical applications in siRNA-based therapies.