Disulfide-crosslinked triblock polyampholyte vesicles as a reducible platform for polyplex-based nucleic acid...

Area of Science:

• Biomaterials Science
• Nanotechnology
• Molecular Biology

Background:

• Polyion complex (PIC) vesicles face challenges in mRNA delivery due to cargo-induced disruption of electrostatic interactions.
• Developing robust carriers is crucial for effective nucleic acid delivery systems.

Purpose of the Study:

• To engineer cargo-tolerant polyion complex vesicles for enhanced mRNA delivery.
• To investigate the efficacy of triblock polyampholyte vesicles (TBPVs) for encapsulating and delivering mRNA.

Main Methods:

• Synthesis of poly(ethylene glycol)-b-poly(L-lysine)-b-poly(aspartic acid) triblock copolymers.
• Assembly of triblock polyampholyte vesicles (TBPVs) and encapsulation of mRNA (free and pre-condensed polyplexes).
• Assessment of vesicle integrity, protein expression (in vitro and in vivo), and stability (disulfide crosslinking).

Main Results:

• TBPVs demonstrated robust assembly independent of cargo charge, maintaining vesicle integrity.
• Encapsulated pre-condensed mRNA polyplexes in TBPVs led to significant protein expression in vitro and in vivo.
• Disulfide crosslinked TBPVs (SS-TBPVs) showed physiological stability and redox-responsive release, enabling hepatic transgene expression.

Conclusions:

• Triblock polyampholyte vesicles offer a versatile and cargo-tolerant platform for polyplex-based nucleic acid delivery.
• SS-TBPVs are a promising system for stable and targeted mRNA delivery, facilitating gene expression.
• This work advances the development of advanced carriers for therapeutic nucleic acid applications.