Localization of human coagulation factor VIII (hFVIII) in transgenic rabbit by FISH-TSA: identification of transgene copy number and transmission to the next generation

Folia Biol (Praha). 2008;54(4):121-4.

Abstract

For chromosomal localization of the hFVIII human transgene in F2 and F3 generation of transgenic rabbits, FISH-TSA was applied. A short cDNA probe (1250 bp) targeted chromosomes 3, 7, 8, 9 and 18 of an F2 male (animal 1-3-8). Two transgenic offspring (F3) revealed signal positions in chromosome 3 and chromosomes 3 and 7, respectively. Sequencing and structure analysis of the rabbit orthologous gene revealed high similarity to its human counterpart. Part of the sequenced cDNA (1310 bp) served as a probe for FISH-TSA analysis. The rabbit gene was localized in the q arm terminus of the X chromosome. This result is in agreement with reciprocal chromosome painting between the rabbit and the human. The presented FISH-TSA method provides strong signals without any interspecies reactivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Chromosome Mapping / methods*
  • Chromosomes, Mammalian*
  • Factor VIII / genetics*
  • Factor VIII / metabolism
  • Female
  • Gene Dosage*
  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • Male
  • Nucleic Acid Amplification Techniques*
  • Rabbits
  • Transgenes / genetics

Substances

  • Factor VIII