Homozygous mice for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. B-ROSA26-EGFP-DTA mice have STOP cassettes preventing expression of diphtheria toxin fragment A (DTA). DTA expression will be activated, if STOP cassettes are removed – resulting in the specific ablation of the cre-expressing cells. ROSA26-EGFP-DTA mice are useful for Cre-inducible deletion of specific groups of cells.
Gene editing strategy
A targeting vector was designed to insert an porcine teschovirus-1 2A (P2A) linked to iCre recombinase gene, downstream of the proge sterone receptor (Pgr) stop codon. The targeting vector was electroporated into C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulted chimeric mice. These mice were maintained on a C57BL/6 background.
(a) The reproductive system of mice, including the uterus, fallopian tubes and ovaries. (b) Mouse mammary glands. The expressions of red fluorescent protein and green fluorescent labeled Pgr (progesterone) of tdTomato were detected by IF. The vertical axis represents mice with different genotypes, and the horizontal axis represents different fluorescent markers. B-Pgr-iCre(Mut/+); CAG-tdTomato (Mut/+) Mice with genotype showed obvious expression of red fluorescent protein of tdTomato.
1. Soyal SM, Mukherjee A, Lee KY, Li J, Li H, DeMayo FJ, Lydon JP. Cre-mediated recombination in cell lineages that express the progesterone receptor. Genesis. 2005 Feb;41(2):58-66. [PubMed: 15682389]
2. Ismail PM, Li J, DeMayo FJ, O’Malley BW, Lydon JP. A novel LacZ reporter mouse reveals complex regulation of the progesterone receptor promoter during mammary gland development. Mol Endocrinol. 2002 Nov;16(11):2475-89. [PubMed: 12403837]
3. Lydon JP, DeMayo FJ, Funk CR, Mani SK, Hughes AR, Montgomery CA Jr, Shyamala G, Conneely OM, O’Malley BW. Mice lacking progesterone receptor exhibit pleiotropic reproductive abnormalities. Genes Dev. 1995 Sep 15;9(18):2266-78. [PubMed: 7557380]
4. Yang CF, Chiang MC, Gray DC, Prabhakaran M, Alvarado M, Juntti SA, Unger EK, Wells JA, Shah NM. Sexually dimorphic neurons in the ventromedial hypothalamus govern mating in both sexes and aggression in males. Cell. 2013 May 9;153(4):896-909. [PubMed: 23663785]
5. Tibbetts TA, Mendoza-Meneses M, O’Malley BW, Conneely OM. Mutual and intercompartmental regulation of estrogen receptor and progesterone receptor expression in the mouse uterus. Biol Reprod. 1998 Nov;59(5):1143-52. [PubMed: 9780321]