Microinjection of targeted embryonic stem cells and establishment of knockout mouse lines for Fmo genes

Methods Mol Biol. 2006:320:329-41.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / enzymology
  • Female
  • Guidelines as Topic
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microinjections
  • Oxygenases / genetics
  • Oxygenases / physiology*
  • Pregnancy
  • Stem Cells / cytology*
  • Stem Cells / enzymology

Substances

  • Oxygenases
  • dimethylaniline monooxygenase (N-oxide forming)