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Related Experiment Videos

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

S Miyagawa1, M Yamada, K Matsunami

  • 1Division of Organ Transplantation, Osaka University Graduate School of Medicine, Yamadaoka, Suita, Osaka 565-0871, Japan. miyagawa@orgtrp.med.osaka-u.ac.jp

Journal of Biochemistry
|May 1, 2001
PubMed
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Synthetically modified human DAF (CD55) gene enhanced protein production in cells and transgenic mice without increasing mRNA levels. This synthetic DAF (sDAF) shows promise for developing high-expression transgenic pigs for xenotransplantation.

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).

Related Experiment Videos

  • 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.