Rapidly Visualizing the Membrane Affinity of Gene Vectors Using Polydiacetylene-Based Allochroic Vesicles

Area of Science:

• Biomaterials Science
• Nanotechnology
• Gene Delivery

Background:

• High-throughput screening is crucial for drug development, but gene carrier screening is often overlooked.
• Gene carriers directly influence drug efficacy and in vivo fate by interacting with physiological barriers and cells.
• Developing efficient and safe gene delivery systems requires understanding carrier-cell interactions.

Purpose of the Study:

• To construct polydiacetylene (PDA) vesicles that mimic cell membranes for detecting gene vector membrane affinity.
• To establish a rapid, visual method for predicting gene vector performance based on membrane interaction.
• To correlate in vitro membrane affinity measurements with in vivo gene transfection efficiency.

Main Methods:

• Synthesized polydiacetylene (PDA) vesicles (approx. 550 nm) with tunable surface potentials by altering phospholipid composition.
• Utilized naked-eye visual color change detection within 5 minutes upon gene vector addition.
• Assessed membrane affinity of commercial gene vectors (Lipofectamine 2000, Entranster-H4000, polyethylenimine) using optimized 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)-PDA vesicles.
• Correlated logarithm of transfection efficiency (pGFP) in L02 and HepG2 cells with logarithm of membrane affinity (log Kb).

Main Results:

• All constructed vesicles exhibited rapid color changes (<5 min) with gene vectors.
• Optimized DMPC-PDA vesicles demonstrated the most sensitive response to tested gene vectors.
• A strong linear correlation was observed between gene vector membrane affinity (log Kb) and transfection efficiency (log pGFP).

Conclusions:

• The DMPC-PDA vesicle system provides a rapid, visual in vitro method for predicting gene vector membrane affinity.
• This method accurately correlates with gene transfection efficiency, aiding in the selection of effective gene delivery vectors.
• The developed strategy offers a universal approach for high-throughput screening of novel carrier materials with high cell affinity.