title: Microinjection of Targeted Embryonic Stem Cells and Establishment of Knockout Mouse Lines for Fmo Genes.
abstract: Methods are described for the injection of mouse embryonic stem cells, in which Fmo genes have been targeted to disrupt gene function, into 3.5-d-old blastocysts and the implantation of these into foster mothers. Successful injection and implantation of blastocysts will produce mice of mixed coat color (the chimera). Also described are methods to establish the success of blastocyst injection and implantation of germ-line transmission of the knockout (KO) mutation. Breeding strategies to produce congenic and isogenic KO mouse lines are outlined. Simple methods for the isolation of tail DNA, the tagging of mice, and record keeping of the line are also given.
authors.affiliation: Department of Biochemistry and Molecular Biology, University College London, London, UK.
authors.forename: Diana
authors.initials: D
authors.lastname: Hernandez
authors.affiliation: Laboratory of Biology, School of Medicine, University of Ioannina, Ioannina, Greece.
authors.forename: Anna N
authors.initials: AN
authors.lastname: Melidoni
authors.affiliation: School of Biological Sciences, Queen Mary, University of London, London, UK.
authors.forename: Ian R
authors.initials: IR
authors.lastname: Phillips
authors.affiliation: Department of Biochemistry and Molecular Biology, University College London, London, UK.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Shephard
keywords: Knockout mice; blastocysts, tail DNA; breeding strategy; mouse colony; mouse embryonic stem cells