Article

Mammalian Genome

, Volume 24, Issue 7, pp 286-294

First online:

Open Access This content is freely available online to anyone, anywhere at any time.

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

  • Edward RyderAffiliated withThe Wellcome Trust Sanger Institute Email author 
  • , Diane GleesonAffiliated withThe Wellcome Trust Sanger Institute
  • , Debarati SethiAffiliated withThe Wellcome Trust Sanger Institute
  • , Sapna VyasAffiliated withThe Wellcome Trust Sanger Institute
  • , Evelina MiklejewskaAffiliated withThe Wellcome Trust Sanger Institute
  • , Priya DalviAffiliated withThe Wellcome Trust Sanger Institute
  • , Bishoy HabibAffiliated withThe Wellcome Trust Sanger Institute
  • , Ross CookAffiliated withThe Wellcome Trust Sanger Institute
  • , Matthew HardyAffiliated withThe Wellcome Trust Sanger Institute
    • , Kalpesh JhaveriAffiliated withThe Wellcome Trust Sanger Institute
    • , Joanna BottomleyAffiliated withThe Wellcome Trust Sanger Institute
    • , Hannah Wardle-JonesAffiliated withThe Wellcome Trust Sanger Institute
    • , James N. BussellAffiliated withThe Wellcome Trust Sanger Institute
    • , Richard HoughtonAffiliated withThe Wellcome Trust Sanger Institute
    • , Jennifer SalisburyAffiliated withThe Wellcome Trust Sanger Institute
    • , William C. SkarnesAffiliated withThe Wellcome Trust Sanger Institute
    • , Sanger Mouse Genetics ProjectAffiliated withThe Wellcome Trust Sanger Institute
    • , Ramiro Ramirez-SolisAffiliated withThe Wellcome Trust Sanger Institute Email author 

Abstract

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.