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	ES Cell Microinjection

The production of chimeric mice by the introduction of embryonic stem (ES) cells into early embryos is an another approach to generating transgenic mice. The technique utilizes a tissue culture system to modify and select ES cell that carries an exogenous DNA of interest. Once derived and characterized, ES cell clones may be transferred into 4-day-old mouse embryos where they will differentiate into adult tissue.

The facility will inject targeted ES cells containing site specific genomic alterations (knockouts) provided by the investigator into 40-50 blastocysts of C57B2/6 mice. When the blastocysts have reexpanded, they are transferred into the uterine horn of a three day pseudopregnant female and allowed to develop to term. Upon weaning 3-4 weeks after birth, chimeric mice are separated and then released to the care of the investigator. These mice can then be bred in order to pass the targeted mutation in the germline.

Difficulties in creating chimeric mice.  

a)     A MAP test is required for any new ES cell lines prior to microinjection.

b)     On the assigned day, the freshly harvested ES cell sample (without feeder cells) must be delivered to transgenic facility in order to proceed with the injection.