B6.Cg-Gt(ROSA)26Sor^{tm2.1(CAG-EGFP, -DTA*G128D)Pjen}/J

Gt(ROSA)26Sor^{tm2.1(CAG-EGFP,-DTA*G128D)Pjen}

targeted mutation 2.1

Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence

Gt(ROSA)26Sor

gene trap ROSA 26

6

Dta, Diphtheria toxin A chain; GFP, Green Fluorescent Protein

5702438

EGFP fluorescence is widespread in the absence of Cre recombinase. Subsequent exposure to Cre recombinase allows expression of attenuated diphtheria toxin (DTA*G128D) resulting in ablation of those Cre expressing cells.

(129S6/SvEvTac x C57BL/6NCrl)F1

The targeting vector iss designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a rox-flanked His3-SV40 transcriptional STOP cassette, a FRT flanked His3-SV40 transcriptional STOP cassette, a Cre-dependent FLEx switch (containing [from 5' to 3'] an inward-facing lox2272 site, a cDNA sequence encoding enhanced green fluorescent protein [eGFP], a rabbit -globin polyA signal, an inward-facing loxP site, a reverse complimentary sequence encoding the tox 176 attenuated diphtheria toxin A chain [G-to-A transition at nt 383 encodes glycine-to-aspartic acid at amino acid 128 (DTA*G128D)], a second inward-facing lox2272 site, a second inward-facing loxP site), a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, and an attB/attP-flanked PGK-FRT Neo-polyA cassette. This entire targeting vector is inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. A PhiC31-expressing plasmid is used to remove the attB/attP-flanked PGK-FRT Neo-polyA cassette and replace it with the recombined attB/attP site (attL). Flp-mediated recombination removed the FRT flanked STOP sequence. Dre-mediated recombination removed the rox-flanked STOP sequence. The resulting allele has a CAG hybrid promoter followed by a variant lox-flanked region containing an eGFP cassette and an inverted attenuated diptheria toxin subunit alpha gene (DTA*G128D). GFP expression is visualized by direct fluorescence in embryos and adult brain.

C57BL/6J

When maintaining a live colony, heterozygous females may be bred to wildtype males from the colony, or to C57BL/6J inbred males. The donating investigator reports that homozygous males appear to be infertile (unknown cause), and heterozygous males have been very sub-fertile.

Heterozygote x Wild-type