Microinjection of targeted embryonic stem cells and establishment of knockout mouse lines for Fmo genes.
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.
Hernandez
Diana
D
Department of Biochemistry and Molecular Biology, University College London, UK.
Melidoni
Anna N
AN
Phillips
lan R
lR
Shephard
Elizabeth A
EA
D000818:Animals; D004622:Embryo, Mammalian / Q000166:cytology* / Q000201:enzymology; D005260:Female; D017408:Guidelines as Topic; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008845:Microinjections; D010105:Oxygenases / Q000235:genetics / Q000502:physiology*; D011247:Pregnancy; D013234:Stem Cells / Q000166:cytology* / Q000201:enzymology
D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)