title: Blastocyst genotyping for quality control of mouse mutant archives: an ethical and economical approach.
abstract: With the advent of modern developmental biology and molecular genetics, the scientific community has generated thousands of newly genetically altered strains of laboratory mice with the aim of elucidating gene function. To this end, a large group of Institutions which form the International Mouse Phenotyping Consortium is generating and phenotyping a knockout mouse strain for each of the ~20,000 protein-coding genes using the mutant ES cell resource produced by the International Knockout Mouse Consortium. These strains are made available to the research community via public repositories, mostly as cryopreserved sperm or embryos. To ensure the quality of this frozen resource there is a requirement that for each strain the frozen sperm/embryos are proven able to produce viable mutant progeny, before the live animal resource is removed from cages. Given the current requirement to generate live pups to demonstrate their mutant genotype, this quality control check necessitates the use and generation of many animals and requires considerable time, cage space, technical and economic resources. Here, we describe a simple and efficient method of genotyping pre-implantation stage blastocysts with significant ethical and economic advantages especially beneficial for current and future large-scale mouse mutagenesis projects.
authors.affiliation: Consiglio Nazionale delle Ricerche (IBCN), CNR-Campus International Development (EMMA-INFRAFRONTIER- IMPC), A. Buzzati-Traverso Campus, Via E. Ramarini 32, 00015, Monterotondo Scalo, Roma, Italy.
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Scavizzi
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Edward
authors.initials: E
authors.lastname: Ryder
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Stuart
authors.initials: S
authors.lastname: Newman
authors.affiliation: Consiglio Nazionale delle Ricerche (IBCN), CNR-Campus International Development (EMMA-INFRAFRONTIER- IMPC), A. Buzzati-Traverso Campus, Via E. Ramarini 32, 00015, Monterotondo Scalo, Roma, Italy.
authors.forename: Marcello
authors.initials: M
authors.lastname: Raspa
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Diane
authors.initials: D
authors.lastname: Gleeson
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Hannah
authors.initials: H
authors.lastname: Wardle-Jones
authors.affiliation: Department of Molecular and Cellular Biology, National Centre for Biotechnology (CNB-CSIC), Campus de Cantoblanco, Darwin 3, 28049, Madrid, Spain.|CIBERER, ISCIII, Madrid, Spain.
authors.forename: Lluis
authors.initials: L
authors.lastname: Montoliu
authors.affiliation: Department of Molecular and Cellular Biology, National Centre for Biotechnology (CNB-CSIC), Campus de Cantoblanco, Darwin 3, 28049, Madrid, Spain.|CIBERER, ISCIII, Madrid, Spain.
authors.forename: Almudena
authors.initials: A
authors.lastname: Fernandez
authors.affiliation: CNRS, TAAM-CDTA UPS44, 3B rue de la Férollerie, CS 20057 45071, Orléans Cedex 2, France.
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Dessain
authors.affiliation: CNRS, TAAM-CDTA UPS44, 3B rue de la Férollerie, CS 20057 45071, Orléans Cedex 2, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Larrigaldie
authors.affiliation: Karolinska Center for Transgene Technologies, Comparative Medicine, Karolinska Institutet, von Eulers väg 4a, 171 77, Stockholm, Sweden.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Khorshidi
authors.affiliation: Biocenter Oulu, University of Oulu, Aapistie 5 A, 90220, Oulu, Finland.
authors.forename: Reetta
authors.initials: R
authors.lastname: Vuolteenaho
authors.affiliation: Biocenter Oulu, University of Oulu, Aapistie 5 A, 90220, Oulu, Finland.
authors.forename: Raija
authors.initials: R
authors.lastname: Soininen
authors.affiliation: ICS France Institut Clinique de la Souris, PHENOMIN, ICS-MCI, CNRS, INSERM, Université de Strasbourg, 1 rue Laurent Fries, 67404, Illkirch, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: André
authors.affiliation: ICS France Institut Clinique de la Souris, PHENOMIN, ICS-MCI, CNRS, INSERM, Université de Strasbourg, 1 rue Laurent Fries, 67404, Illkirch, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Jacquot
authors.affiliation: Institute of Experimental Genetics, Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
authors.forename: Yi
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Institute of Experimental Genetics, Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
authors.forename: Martin Hrabe
authors.initials: MH
authors.lastname: de Angelis
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
authors.forename: Ramiro
authors.initials: R
authors.lastname: Ramirez-Solis
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK. bd2@sanger.ac.uk.
authors.forename: Brendan
authors.initials: B
authors.lastname: Doe
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D005838:Genotype; D051379:Mice; D011786:Quality Control
keywords: 3R’s; Cryopreservation; Mouse; Network of repositories; PCR; Quality control (QC)