Production of transgenic tilapia with Brockmann bodies secreting [desThrB30] human insulin

Bill Pohajdak, Marc Mansour, Olga Hrytsenko, J. Michael Conlon, L. Clayton Dymond, James R. Wright

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

Background. Tilapia are commercially important tropical fish which, like many teleosts, have anatomically discrete islet organs called Brockmann bodies. When transplanted into diabetic nude mice, tilapia islets provide long-term normoglycemia and mammalian-like glucose tolerance profiles. Methods. Using site-directed mutagenesis and linker ligation we have "humanized" the tilapia insulin gene so that it codes for [desThrB30] human insulin while maintaining the tilapia regulatory sequences. Following microinjection into fertilized eggs, we screened DNA isolated from whole fry shortly after hatching by PCR. Positive fish were grown to sexual maturity and mated to wild-types and positive F1's were further characterized. Results. Human insulin was detected in both serum and in the clusters of β cells scattered throughout the Brockmann bodies. Surrounding non-β cells as well as other tissues were negative indicating β cell specific expression. Purification and sequencing of both A-and B-chains verified that the insulin was properly processed and humanized. Conclusions. After extensive characterization, transgenic tilapia could become a suitable, inexpensive source of islet tissue that can be easily mass-produced for clinical islet xenotransplantation. Because tilapia islets are exceedingly resistant to hypoxia by mammalian standards, transgenic tilapia islets should be ideal for xenotransplantation using immunoisolation techniques.

Original languageEnglish
Pages (from-to)313-323
Number of pages11
JournalTransgenic Research
Volume13
Issue number4
DOIs
Publication statusPublished - Aug 2004

Keywords

  • diabetes
  • islets
  • teleosts
  • transgenic fish
  • xenotransplantation

ASJC Scopus subject areas

  • Biotechnology
  • Animal Science and Zoology
  • Agronomy and Crop Science
  • Genetics

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