[Establishment of a high efficient human coagulation factor VIII eukaryotic expression system using lentiviral...

Area of Science:

• Biotechnology
• Gene Therapy
• Molecular Biology

Context:

• Hemophilia A is a genetic disorder caused by deficiency in coagulation factor VIII (FVIII).
• Developing efficient and safe expression systems for FVIII is crucial for therapeutic applications.
• Lentiviral vectors offer a promising platform for stable gene delivery and expression in mammalian cells.

Purpose:

• To construct a high-efficiency eukaryotic stable expression system for human coagulation factor VIII (FVIII) using a lentiviral vector.
• To assess the biosafety of the established FVIII expression system.

Summary:

• A lentiviral transfer plasmid encoding human B-domain-deleted FVIII (BDDhFVIII) was successfully constructed.
• Infection of CHO cells with the lentivirus resulted in efficient BDDhFVIII expression, with high antigen and activity levels detected.
• RT-PCR confirmed BDDhFVIII mRNA transcription, and biosafety evaluation showed no detectable gag gene or infectious virus in the supernatant.

Impact:

• Establishes a robust and safe lentiviral expression system for human FVIII in CHO cells.
• Provides a foundation for potential large-scale production of therapeutic FVIII.
• Demonstrates the potential of lentiviral vectors for efficient gene therapy applications in coagulation disorders.