Metaphase FISHing of transgenic mice recommended: FISH and SKY define BAC-mediated balanced translocation

Brett S. Abrahams, Annabelle C.O. Chong, Maimun Nisha, Danielle Milette, David A. Brewster, Melissa L. Berry, Farkhad Muratkhodjaev, Sabine Mai, Evica Rajcan-Separovic, Elizabeth M. Simpson

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

The evolving trend to use larger transgenes and their associated increased chance of unexpected genetic events mandates more careful characterization of transgenic mice. In characterizing our five new mouse strains transgenic for the BAC, bEMS4, we have identified the highest copy number reported to date: the stable incorporation of -40 copies of a 194-kb expressed transgene in a single insertion site. We caution, however, that standard molecular techniques failed to identify a balanced translocation in another strain, and an inappropriate site of insertion in a third. Molecular cytogenetic analysis using metaphase FISH was the minimum level of characterization needed to reveal these unexpected genetic events. In addition, we combined FISH and SKY to identify the transgene at the breakpoints of the balanced translocation, t(3;9). This is the first description of a BAC-mediated chromosomal rearrangement and the first application of SKY to identify transgene-induced chromosomal rearrangements.

Original languageEnglish (US)
Pages (from-to)134-141
Number of pages8
JournalGenesis (United States)
Volume36
Issue number3
DOIs
StatePublished - Jul 1 2003
Externally publishedYes

Keywords

  • Bacterial artificial chromosome
  • Balanced translocation
  • Breakpoints
  • FISH
  • Nr2e1
  • Orphan receptor
  • SKY
  • Tailless
  • Transgenic mice

ASJC Scopus subject areas

  • Genetics
  • Endocrinology
  • Cell Biology

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