Expression trapping: identification of novel genes expressed in hematopoietic and endothelial lineages by gene trapping in ES cells.
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.
Stanford
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WL
Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Caruana
G
G
Vallis
K A
KA
Inamdar
M
M
Hidaka
M
M
Bautch
V L
VL
Bernstein
A
A
D000818:Animals; D019070:Cell Lineage; D002999:Clone Cells / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D005796:Genes / Q000235:genetics*; D017930:Genes, Reporter; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D007763:Lac Operon / Q000235:genetics; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D016254:Mutagenesis, Insertional / Q000379:methods*; D013234:Stem Cells / Q000378:metabolism