A universal single-plasmid, single-promoter system for efficient protein display and RNA encapsulation in MS2...

Area of Science:

• Biotechnology
• Molecular Biology
• Virology

Background:

• Bacteriophage MS2 virus-like particles (VLPs) show potential in biomedical fields.
• Current methods face challenges in protein display, RNA encapsulation, and purification.

Purpose of the Study:

• To develop a streamlined system for producing multifunctional MS2 VLPs.
• To enable modular assembly of VLPs with enhanced features.

Main Methods:

• Developed a universal single-plasmid single-promoter (USS) system.
• Utilized PstI and KpnI sites for modular VLP assembly.
• Employed His-tag affinity chromatography for purification.

Main Results:

• Efficient display of large proteins (EGFP, Nanobody N16) and RNA encapsulation.
• Achieved rapid one-step purification, avoiding ultracentrifugation.
• Demonstrated VLP thermal stability and nuclease resistance.
• Validated diagnostic applications (RT-qPCR controls) and dual-delivery functions (gene silencing and protein binding).

Conclusions:

• The USS system offers an efficient, scalable, and simplified approach to VLP production.
• This platform facilitates applications ranging from molecular diagnostics to combination therapies.