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Transgenic Res. 2015 Oct;24(5):921-7. doi: 10.1007/s11248-015-9897-1. Epub 2015 Jul 16.

Blastocyst genotyping for quality control of mouse mutant archives: an ethical and economical approach.

Author information

1
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.
2
Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
3
Department of Molecular and Cellular Biology, National Centre for Biotechnology (CNB-CSIC), Campus de Cantoblanco, Darwin 3, 28049, Madrid, Spain.
4
CIBERER, ISCIII, Madrid, Spain.
5
CNRS, TAAM-CDTA UPS44, 3B rue de la Férollerie, CS 20057 45071, Orléans Cedex 2, France.
6
Karolinska Center for Transgene Technologies, Comparative Medicine, Karolinska Institutet, von Eulers väg 4a, 171 77, Stockholm, Sweden.
7
Biocenter Oulu, University of Oulu, Aapistie 5 A, 90220, Oulu, Finland.
8
ICS France Institut Clinique de la Souris, PHENOMIN, ICS-MCI, CNRS, INSERM, Université de Strasbourg, 1 rue Laurent Fries, 67404, Illkirch, France.
9
Institute of Experimental Genetics, Helmholtz Zentrum München-German Research Center for Environmental Health (GmbH), Neuherberg, Germany.
10
Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK. bd2@sanger.ac.uk.

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.

KEYWORDS:

3R’s; Cryopreservation; Mouse; Network of repositories; PCR; Quality control (QC)

PMID:
26178246
PMCID:
PMC4569667
DOI:
10.1007/s11248-015-9897-1
[Indexed for MEDLINE]
Free PMC Article

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