Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

Weisheng V. Chen; Jeffrey Delrow; Philip D. Corrin; Jason P. Frazier; Philippe Soriano

doi:10.1038/ng1306

Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

Weisheng V. Chen, Jeffrey Delrow, Philip D. Corrin, Jason P. Frazier, Philippe Soriano

Research output: Contribution to journal › Article › peer-review

99 Scopus citations

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.

Original language	English
Pages (from-to)	304-312
Number of pages	9
Journal	Nature Genetics
Volume	36
Issue number	3
DOIs	https://doi.org/10.1038/ng1306
State	Published - Mar 2004
Externally published	Yes

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title = "Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis",

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.",

author = "Chen, {Weisheng V.} and Jeffrey Delrow and Corrin, {Philip D.} and Frazier, {Jason P.} and Philippe Soriano",

note = "Funding Information: We thank C. Neal for expert assistance in microarray fabrication, M. Aronszajn for biocomputing and expert sequence analysis, and S. Tapscott, S. Parkhurst and our laboratory colleagues for critical reading of the manuscript. This work was supported by a grant from the US National Institutes of Child Health and Human Development to P.S.",

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AU - Corrin, Philip D.

AU - Frazier, Jason P.

AU - Soriano, Philippe

N1 - Funding Information: We thank C. Neal for expert assistance in microarray fabrication, M. Aronszajn for biocomputing and expert sequence analysis, and S. Tapscott, S. Parkhurst and our laboratory colleagues for critical reading of the manuscript. This work was supported by a grant from the US National Institutes of Child Health and Human Development to P.S.

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Identification and validation of PDGF transcriptional targets by microarray-coupled gene-trap mutagenesis

Abstract

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