Homozygous ROSA26 GNZ knock-in mice have widespread expression of a nuclear-localized green fluorescent protein/beta-galactosidase fusion protein (GFP-NLS-lacZ or GNZ) once an upstream loxP-flanked STOP sequence is removed. When bred to cre expressing mice, the resulting GNZ fusion protein expression in the offspring allows for enhanced (single cell level) visualization.
Jonathan A Epstein, University of Pennsylvania
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Homozygous ROSA26 GNZ knock-in mice are viable and fertile, with a nuclear-localized green fluorescent protein/beta-galactosidase fusion protein (GFP-NLS-lacZ or GNZ) inserted into the Gt(ROSA)26Sor (ROSA26) locus. Widespread expression of GNZ is blocked by an upstream loxP-flanked STOP sequence (in the absence of Cre recombinase, no expressed GFP or beta-galactosidase activity is observed in GNZ embryos(E9.5-18.5)). When bred to cre expressing mice, offspring will have the STOP sequence deleted in tissues where Cre recombinase is present. The resulting GNZ fusion protein expression allows for enhanced (single cell level) visualization / resolution. The donating investigator reports that Cre recombinase activity can be visualized by direct GFP fluorescence, but the high resolution nuclear staining of GNZ may be best visualized by immunostaining for either GFP or beta-galactosidase. These ROSA26 GNZ mice are useful as a Cre reporter strain; expressing both GFP and beta-galactosidase following Cre-mediated recombination.
A targeting vector was designed with a loxP-flanked STOP cassette (PGK-Neo-polyA followed by transcriptional stop sequence) just upstream of GFP-NLS-lacZ (or GNZ; containing a "humanized" green fluorescent protein (GFP-H or gfph), SV40 Large-T antigen nuclear localization signal (NLS), and beta-galactosidase (lacZ) sequence). This entire construct was inserted into the Gt(ROSA)26Sor locus via electroporation of (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were microinjected into recipient blastocysts and chimeric mice were bred to generate the mutant strain. These GNZ mice were maintained on a mixed C57BL/6J;129 genetic background prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | Mice express a nuclear-localized GFP/beta-galactosidase fusion protein (GFP-NLS-lacZ or GNZ) once an upstream loxP-flanked STOP sequence is removed via a tissue-specific cre recombinase. |
Allele Name | targeted mutation 1, Jonathan A Epstein |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | GFP-NLS-lacZ; GNZ; ROSA26-GNZ; ROSA-nGFP |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Mice express a nuclear-localized GFP/beta-galactosidase fusion protein (GFP-NLS-lacZ or GNZ) once an upstream loxP-flanked STOP sequence is removed via a tissue-specific cre recombinase. |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 6 |
Molecular Note | A targeting vector was designed with a loxP-flanked STOP cassette (PGK-Neo-polyA followed by transcriptional stop sequence) just upstream of a green fluorescent protein/beta-galactosidase fusion protein sequence with SV40 nuclear localization signal (GFP-NLS-lacZ or GNZ). This entire construct was inserted into the Gt(ROSA)26Sor locus via electroporation. |
Mutations Made By | Jonathan Epstein, University of Pennsylvania |
When maintaining a live colony, homozygous mice may be bred.
When using the B6;129-Gt(ROSA)26Sortm1Joe/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #008516 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Gt(ROSA)26Sor<tm1Joe> |
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