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Systematic generation of in vivo G protein-coupled receptor mutants in the rat

Boxtel van, R. and Vroling, B. and Toonen, P.W. and Nijman, I.J. and Roekel van, H. and Verheul, M. and Baakman, C. and Guryev, V. and Vriend, G. and Cuppen, E. (2010) Systematic generation of in vivo G protein-coupled receptor mutants in the rat. Pharmacogenomics J. ISSN 1470-269X.

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Official URL: http://dx.doi.org/10.1038/tpj.2010.44

Abstract

G-protein-coupled receptors (GPCRs) constitute a large family of cell surface receptors that are involved in a wide range of physiological and pathological processes, and are targets for many therapeutic interventions. However, genetic models in the rat, one of the most widely used model organisms in physiological and pharmacological research, are largely lacking. Here, we applied N-ethyl-N-nitrosourea (ENU)-driven target-selected mutagenesis to generate an in vivo GPCR mutant collection in the rat. A pre-selected panel of 250 human GPCR homologs was screened for mutations in 813 rats, resulting in the identification of 131 non-synonymous mutations. From these, seven novel potential rat gene knockouts were established as well as 45 lines carrying missense mutations in various genes associated with or involved in human diseases. We provide extensive in silico modeling results of the missense mutations and show experimental data, suggesting loss-of-function phenotypes for several models, including Mc4r and Lpar1. Taken together, the approach used resulted not only in a set of novel gene knockouts, but also in allelic series of more subtle amino acid variants, similar as commonly observed in human disease. The mutants presented here may greatly benefit studies to understand specific GPCR function and support the development of novel therapeutic strategies.The Pharmacogenomics Journal advance online publication, 8 June 2010; doi:10.1038/tpj.2010.44.

Item Type:Article
Institutes:Hubrecht Instituut
ID Code:9528
Deposited On:11 Feb 2011 01:00
Last Modified:07 Sep 2011 17:01

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