title: Molecular characterization of mutant mouse strains generated from the EUCOMM/KOMP-CSD ES cell resource.
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.
authors.affiliation: The Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK. er1@sanger.ac.uk
authors.forename: Edward
authors.initials: E
authors.lastname: Ryder
authors.forename: Diane
authors.initials: D
authors.lastname: Gleeson
authors.forename: Debarati
authors.initials: D
authors.lastname: Sethi
authors.forename: Sapna
authors.initials: S
authors.lastname: Vyas
authors.forename: Evelina
authors.initials: E
authors.lastname: Miklejewska
authors.forename: Priya
authors.initials: P
authors.lastname: Dalvi
authors.forename: Bishoy
authors.initials: B
authors.lastname: Habib
authors.forename: Ross
authors.initials: R
authors.lastname: Cook
authors.forename: Matthew
authors.initials: M
authors.lastname: Hardy
authors.forename: Kalpesh
authors.initials: K
authors.lastname: Jhaveri
authors.forename: Joanna
authors.initials: J
authors.lastname: Bottomley
authors.forename: Hannah
authors.initials: H
authors.lastname: Wardle-Jones
authors.forename: James N
authors.initials: JN
authors.lastname: Bussell
authors.forename: Richard
authors.initials: R
authors.lastname: Houghton
authors.forename: Jennifer
authors.initials: J
authors.lastname: Salisbury
authors.forename: William C
authors.initials: WC
authors.lastname: Skarnes
authors.forename: Ramiro
authors.initials: R
authors.lastname: Ramirez-Solis
mesh_terms: D000818:Animals; D001947:Breeding; D053595:Embryonic Stem Cells / Q000166:cytology*; D005854:Germ Cells / Q000166:cytology*; D051379:Mice; D008817:Mice, Mutant Strains / Q000235:genetics*; D011786:Quality Control
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