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Publication : Large-scale, saturating insertional mutagenesis of the mouse genome.

First Author  Gragerov A Year  2007
Journal  Proc Natl Acad Sci U S A Volume  104
Issue  36 Pages  14406-11
PubMed ID  17720809 Mgi Jnum  J:124855
Mgi Id  MGI:3722704 Doi  10.1073/pnas.0700608104
Citation  Gragerov A, et al. (2007) Large-scale, saturating insertional mutagenesis of the mouse genome. Proc Natl Acad Sci U S A 104(36):14406-11
abstractText  We describe the construction of a large-scale, orderly assembly of mutant ES cells, generated with retroviral insertions and having mutational coverage in >90% of mouse genes. We also describe a method for isolating ES cell clones with mutations in specific genes of interest from this library. This approach, which combines saturating random mutagenesis with targeted selection of mutations in the genes of interest, was successfully applied to the gene families of G protein-coupled receptors (GPCRs) and nuclear receptors. Mutant mouse strains in 60 different GPCRs were generated. Applicability of the technique for the GPCR genes, which on average represent fairly small targets for insertional mutagenesis, indicates the general utility of our approach for the rest of the genome. The method also allows for increased scale and automation for the large-scale production of mutant mice, which could substantially expedite the functional characterization of the mouse genome.
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