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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4622-4631
Expression Trapping: Identification of Novel Genes Expressed in
Hematopoietic and Endothelial Lineages by Gene Trapping in ES Cells
William L. Stanford,
Georgina Caruana,
Katherine A. Vallis,
Maneesha Inamdar,
Michihiro Hidaka,
Victoria L. Bautch, and
Alan Bernstein
From the Program in Molecular Biology and Cancer, Samuel Lunenfeld
Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada; the
Ontario Cancer Institute, Toronto, Ontario, Canada; the Department of
Molecular and Medical Genetics, University of Toronto, Toronto,
Ontario, Canada; and the Department of Biology, University of North
Carolina, Chapel Hill, Chapel Hill, NC.
We have developed a large-scale, expression-based gene trap strategy
to perform genome-wide functional analysis of the murine hematopoietic
and vascular systems. Using two different gene trap vectors, we have
isolated embryonic stem (ES) cell clones containing lacZ
reporter gene insertions in genes expressed in blood island and
vascular cells, muscle, stromal cells, and unknown cell types. Of 79 clones demonstrating specific expression patterns, 49% and 16% were
preferentially expressed in blood islands and/or the vasculature, respectively. The majority of ES clones that expressed lacZ in blood islands also expressed lacZ upon
differentiation into hematopoietic cells on OP9 stromal layers.
Importantly, the in vivo expression of the lacZ fusion products
accurately recapitulated the observed in vitro expression patterns.
Expression and sequence analysis of representative clones suggest that
this approach will be useful for identifying and mutating novel genes
expressed in the developing hematopoietic and vascular systems.
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