Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis.
We developed a versatile, high-throughput genetic screening strategy by coupling gene mutagenesis and expression profiling technologies. Using a retroviral gene-trap vector optimized for efficient mutagenesis and cloning, we randomly disrupted genes in mouse embryonic stem (ES) cells and amplified them to construct a cDNA microarray. With this gene-trap array, we show that transcriptional target genes of platelet-derived growth factor (PDGF) can be efficiently and reliably identified in physiologically relevant cells and are immediately accessible to genetic studies to determine their in vivo roles and relative contributions to PDGF-regulated developmental processes. The same platform can be used to search for genes of specific biological relevance in a broad array of experimental settings, providing a fast track from gene identification to functional validation.
Program in Developmental Biology, Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Weisheng V
WV
Chen
Jeffrey
J
Delrow
Philip D
PD
Corrin
Jason P
JP
Frazier
Philippe
P
Soriano
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D051379:Mice; D016296:Mutagenesis; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism
D010982:Platelet-Derived Growth Factor