Molecular Characterization of Mutant Mouse Strains Generated from the EUCOMM/KOMP-CSD ES Cell Resource

• Published in: Mammalian genome Vol. 24; no. 7-8; pp. 286 - 294
• Main Authors: Ryder, Edward, Gleeson, Diane, Sethi, Debarati, Vyas, Sapna, Miklejewska, Evelina, Dalvi, Priya, Habib, Bishoy, Cook, Ross, Hardy, Matthew, Jhaveri, Kalpesh, Bottomley, Joanna, Wardle-Jones, Hannah, Bussell, James N, Houghton, Richard, Salisbury, Jennifer, Skarnes, William C, Ramirez-Solis, Ramiro
• Format: Journal Article
• Language: English
• Published: Boston Springer-Verlag 01.08.2013; Springer US; Springer Nature B.V
• Subjects: alleles; Animal Genetics and Genomics; Animals; Biomedical and Life Sciences; Breeding; Cell Biology; cytology; Embryonic Stem Cells; Embryonic Stem Cells - cytology; genetics; germ cells; Germ Cells - cytology; Human Genetics; Life Sciences; Mice; Mice, Mutant Strains; Mice, Mutant Strains - genetics; mutants; mutation; phenotype; polymerase chain reaction; Quality Control; screening; Quality Control Method; International Mouse Phenotyping Consortium; Embryonic Stem Cell; Embryonic Stem Cell Clone; loxP Site
• ISSN: 0938-8990; 1432-1777; 1432-1777
• DOI: 10.1007/s00335-013-9467-x

The Sanger Mouse Genetics Project generates knockout mice strains using the EUCOMM/KOMP-CSD embryonic stem (ES) cell collection and characterizes the consequences of the mutations using a high-throughput primary phenotyping screen. Upon achieving germline transmission, new strains are subject to a panel of quality control (QC) PCR- and qPCR-based assays to confirm the correct targeting, cassette structure, and the presence of the 3′ LoxP site (required for the potential conditionality of the allele). We report that over 86 % of the 731 strains studied showed the correct targeting and cassette structure, of which 97 % retained the 3′ LoxP site. We discuss the characteristics of the lines that failed QC and postulate that the majority of these may be due to mixed ES cell populations which were not detectable with the original screening techniques employed when creating the ES cell resource.