Dissemin is shutting down on January 1st, 2025

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Springer, Mammalian Genome, 7-8(24), p. 286-294, 2013

DOI: 10.1007/s00335-013-9467-x

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

Journal article published in 2013 by Edward Ryder, Debarati Sethi, Sapna Vyas, Evelina Miklejewska, Kalpesh Jhaveri, Hannah Wardle-Jones, Richard Houghton, Jennifer Salisbury, William C. Skarnes, Diane Gleeson, Ramiro Ramirez-Solis, Priya Dalvi, Bishoy Habib, Ross Cook, Joanna Bottomley and other authors.
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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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.