Co-delivery of genes can be confounded by bicistronic vector design.

Area of Science:

• Biotechnology
• Molecular Biology
• Nanotechnology

Background:

• Efficient co-delivery of multiple genes into the same cell is crucial for various biological applications.
• Current strategies primarily focus on nanocarrier systems, often overlooking the impact of gene payload design.

Purpose of the Study:

• To investigate the influence of different gene payload designs on nanocarrier-mediated co-delivery efficiency.
• To compare the co-expression rates achieved using monocistronic versus bicistronic nucleic acid (NA) designs.

Main Methods:

• Co-delivery of two individual monocistronic NAs using nanocarriers.
• Co-delivery of a single bicistronic NA containing two genes separated by a 2A self-cleavage site via nanocarriers.
• Quantification of co-expressing cells for each delivery strategy.

Main Results:

• The monocistronic design, involving two separate NAs, resulted in a significantly higher percentage of co-expressing cells compared to the bicistronic design.
• Achieving predictable co-expression with the bicistronic NA design proved challenging.

Conclusions:

• Payload design significantly impacts the efficiency of gene co-delivery mediated by nanocarriers.
• The monocistronic approach is more effective for achieving higher co-expression rates in the same cell.
• These findings provide guidance for selecting optimal gene co-delivery methodologies based on specific application requirements.