Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis
- 15 February 2004
- journal article
- technical report
- Published by Springer Nature in Nature Genetics
- Vol. 36 (3) , 304-312
- https://doi.org/10.1038/ng1306
Abstract
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.Keywords
This publication has 42 references indexed in Scilit:
- Evolutionary Divergence of Platelet-Derived Growth Factor Alpha Receptor Signaling MechanismsMolecular and Cellular Biology, 2003
- Initial sequencing and comparative analysis of the mouse genomeNature, 2002
- Alternative pre-mRNA splicing and proteome expansion in metazoansNature, 2002
- βIV-spectrin regulates sodium channel clustering through ankyrin-G at axon initial segments and nodes of RanvierThe Journal of cell biology, 2002
- Initial sequencing and analysis of the human genomeNature, 2001
- Efficient Gene Trap Screening for Novel Developmental Genes Using IRESβgeo Vector andin VitroPreselectionExperimental Cell Research, 1998
- Induction of Inhibitory Smad6 and Smad7 mRNA by TGF-β Family MembersBiochemical and Biophysical Research Communications, 1998
- Selective disruption of genes expressed in totipotent embryonal stem cells.Genes & Development, 1992
- Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice.Genes & Development, 1991
- PRIMARY RESPONSE GENES INDUCED BY GROWTH FACTORS AND TUMOR PROMOTERSAnnual Review of Biochemistry, 1991