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Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

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Published on: March 1, 2013

Stimuli-Responsive Polypeptide Nanoparticles for Enhanced DNA Delivery.

Olga Korovkina¹, Dmitry Polyakov², Viktor Korzhikov-Vlakh^1,3

¹Institute of Chemistry, Saint-Petersburg State University, Universitetsky pr. 26, 198504 St. Petersburg, Russia.

Molecules (Basel, Switzerland)

|December 11, 2022

Summary

New pH- and redox-responsive polypeptide nanoparticles offer efficient gene therapy delivery. These systems stabilize in the body and release DNA inside cells, achieving 70% transfection efficacy in HEK 293 cells.

Keywords:

cross-linked nanoparticles gene delivery pH and redox-responsive delivery systems polypeptides

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Area of Science:

Biomedicinal Chemistry
Polymer Chemistry
Gene Therapy Delivery

Background:

Developing effective non-viral gene delivery systems remains a significant challenge in biomedicinal chemistry.
Existing systems often struggle with stability and controlled intracellular release.

Purpose of the Study:

To synthesize and characterize novel pH- and redox-responsive amphiphilic polypeptides for enhanced intracellular DNA delivery.
To investigate the self-assembly, stability, and degradation properties of these polypeptide-based nanoparticles.

Main Methods:

Synthesis of polypeptides via N-carboxyanhydride copolymerization and post-polymerization modification.
Characterization of nanoparticle size, surface charge, and stability under various conditions.
Assessment of intracellular DNA release triggered by glutathione and evaluation of gene transfection efficacy in HEK 293 cells.

Main Results:

Polypeptides containing histidine (pH-sensitivity) and cysteine (redox-sensitivity) were successfully synthesized.
Cross-linked nanoparticles showed enhanced stability, compacting from 200-250 nm to 55-100 nm.
Glutathione triggered nanoparticle degradation and nucleic acid release, with optimal DNA delivery at a polypeptide/pDNA ratio of 100, achieving 70% GFP expression.

Conclusions:

The developed polypeptide nanoparticles are promising non-viral vectors for gene therapy.
Their pH and redox responsiveness enable controlled intracellular DNA release and efficient gene transfection.
These systems demonstrate potential for overcoming current gene delivery challenges.