Submitted on November 23, 2005
Accepted on May 10, 2006
N-Ethyl-N-Nitrosurea-Based Generation of Mouse Models for Mutant G-Protein-Coupled Receptors
Johannes Grosse1, Patrick Tarnow2, Holger Rompler3, Boris Schneider1, Reinhard Sedlmeier1, Ulrike Huffstadt1, Dirk Korthaus1, Michael Nehls1, Sigrid Wattler1, Torsten Schoneberg3, Heike Biebermann2, and Martin Augustin1*
1 Ingenium Pharmaceuticals AG, Martinsried, Germany
2 Institute of Pediatric Endocrinology, Humboldt University, Charite Campus Virchow Klinikum, Berlin, Germany
3 Institute of Biochemistry, Molecular Biochemistry, Medical Faculty, University of Leipzig, Leipzig, Germany
* To whom correspondence should be addressed. E-mail: martin.augustin{at}ingenium-ag.com.
Chemical random mutagenesis techniques with the germ line supermutagen N-ethyl-N-nitrosurea (ENU) have been established to provide comprehensive collections of mouse models, which were then mined and analyzed in phenotype-driven studies. Here, we applied ENU-mutagenesis in a high-throughput fashion for a gene-driven identification of new mutations. Selected members of the large superfamily of G-protein-coupled receptors (GPCR), melanocortin type 3 (Mc3r) and type 4 (Mc4r) receptors and the orphan chemoattractant receptor GPR33, were used as model targets to prove the feasibility of this approach. Parallel archives of DNA and sperm from mice mutagenized with ENU were screened for mutations in these GPCR and in vitro assays served as a pre-selection step before in vitro fertilization (IVF) was performed to generate the appropriate mouse model. For example, mouse models for inherited obesity were established by selecting fully or partially inactivating mutations in Mc4r. Our technology described herein has the potential to provide mouse models for a GPCR dysfunction of choice within less than four months and can be extended to other gene classes of interest.
