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

TitleAn induction gene trap screen in embryonic stem cells: Identification of genes that respond to retinoic acid in vitro.
Publication TypeJournal Article
Year of Publication1996
AuthorsForrester LM, Nagy A, Sam M, Watt A, Stevenson L, Bernstein A, Joyner AL, Wurst W
JournalProc Natl Acad Sci U S A
Volume93
Issue4
Pagination1677-82
Date Published1996 Feb 20
ISSN0027-8424
KeywordsAnimals, Cell Lineage, Cells, Cultured, Chimera, Cloning, Molecular, Coculture Techniques, Embryo, Mammalian, Embryonic and Fetal Development, Female, Fibroblasts, Gene Expression Regulation, Developmental, Gene Targeting, Genes, Reporter, Genetic Vectors, Male, Mice, Mice, Transgenic, Recombinant Fusion Proteins, Signal Transduction, Stem Cells, Tretinoin
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.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID8643689
PubMed Central IDPMC40001