A synthetic dDAF (CD55) gene based on optimal codon usage for transgenic animals

Area of Science:

• Biotechnology
• Xenotransplantation research
• Molecular biology

Background:

• The human DAF (CD55) gene is crucial for regulating complement activation.
• Optimizing gene expression is key for successful xenotransplantation strategies.
• Codon adaptation can enhance protein production in heterologous systems.

Purpose of the Study:

• To synthesize a human DAF (CD55) gene with codon optimization for mammalian expression.
• To evaluate the expression levels of synthetic DAF (sDAF) in various cell types and in vivo.
• To assess the potential of sDAF for generating high-expression transgenic pigs.

Main Methods:

• Gene synthesis and codon optimization of human DAF (CD55).
• Expression analysis in Chinese hamster ovarian (CHO) cells and swine endothelial cells (SEC).
• Assessment of sDAF expression in transgenic mice and various organs.

Main Results:

• Synthetic DAF (sDAF) demonstrated significantly increased protein production compared to wild-type DAF (wtDAF) and Delta1wtDAF.
• No detectable increase in mRNA levels was observed with sDAF expression.
• sDAF exhibited higher expression in multiple organs of transgenic mice, particularly the pancreas, and in SEC.

Conclusions:

• Codon-optimized human DAF (sDAF) enhances protein expression in vitro and in vivo.
• sDAF shows high expression potential in swine cells and transgenic mice.
• sDAF is a promising candidate for developing transgenic pigs with enhanced DAF expression for xenotransplantation.