Expert Reviews in Molecular Medicine: http://www-ermm.cbcu.cam.ac.uk
Accession information: (01)00297-6h.htm (shortcode: fig002jme); 11 May 2001
Two methods to produce transgenic mice
Jean C. Manson and Nadia L. Tuzi
Author contact details
Figure 2. Two methods to produce transgenic mice. (a) Standard transgenic approach. DNA containing the transgene is microinjected into the male pronucleus of a murine oocyte, which is then transferred to a 0.5-day pseudopregnant recipient mouse. Offspring are screened for the presence of the transgene. (b) Gene-targeted transgenic approach. Isogenic DNA [to the embryonic stem (ES) cells being targeted] possessing the transgene is introduced into the ES cells by, for example, electroporation. Drug selection is used and surviving colonies are screened for the presence of the transgene. Characterised targeted ES cells are microinjected into 3.5-day mouse blastocysts and transferred to 2.5-day pseudopregnant recipient mice. The incorporation of targeted ES cells into the offspring is determined by coat colour – i.e. chimaeric mice are produced that possess coat colour of the mouse that donated blastocysts and that of the mouse from which the ES cells were derived. Chimaeric mice are mated to determine whether the targeted ES cells have contributed to the germline. Germline offspring are screened for the presence of the transgene and mated to establish the transgenic line (not shown) (fig002jme).