An induction gene trap screen in embryonic stem cells: Identification of genes that respond to retinoic acid in vitro

Lesley M. Forrester, Andras Nagy, Mehran Sam, Alistair Watt, Lois Stevenson, Alan Bernstein, Alexandra L. Joyner, Wolfgang Wurst

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108 Citations (Scopus)

Abstract

We have developed a novel induction gene trap approach that preselects in vitro for integrations into genes that lie downstream of receptor/ligand- mediated signaling pathways. Using this approach, we have identified 20 gene trap integrations in embryonic stem cells, 9 of which were induced and 11 of which were repressed after exposure to exogenous retinoic acid (RA). All but one of these integrations showed unique spatially restricted or tissue- specific patterns of expression between 8.5 and 11.5 days of embryogenesis. Interestingly, expression was observed in tissues that are affected by alterations in RA levels during embryogenesis. Sequence analysis of fusion transcripts from six integrations revealed five novel gene sequences and the previously identified protooncogene c-fyn. To date, germ-line transmission and breeding has uncovered one homozygous embryonic lethal and three homozygous viable insertions. These studies demonstrate the potential of this induction gene trap approach for identifying and mutating genes downstream of signal transduction pathways.

Original languageEnglish
Pages (from-to)1677-1682
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number4
DOIs
Publication statusPublished - 20 Feb 1996
Externally publishedYes

Keywords

  • genetic screen
  • induction trapping

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