| Allele Name | targeted mutation 1, Simon Rumpel |
| Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter) |
| Allele Synonym(s) | R26 PA-GFP::NLS |
| Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
| Gene Synonym(s) | |
| Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
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| Expressed Gene | GFP, Green Fluorescent Protein, |
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| Site of Expression | When bred to mice that express Cre recombinase, photoactivatable GFP will be expressed in tissues where promoters driving Cre recombinase are expressed. |
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| Strain of Origin | (C57BL/6 x 129P2/OlaHsd)F1 |
| Chromosome | 6 |
| General Note | After removal of the loxP-flanked cassette via cre-mediated recombination, PA-GFP is expressed in all cells where Cre recombinase is expressed. PA-GFP::NLS is enriched in the nucleus, but can be also detected in the cytoplasm of neuronal cells. PA-GFP shows very weak fluorescence at 517nm when excited at 488nm before photoactivation. Upon photoactivation with light at ~400nm PA-GFP is converted into its fluorescent form. This form shows strong emission at 517nm when excited at 488nm and allows the conditional fluorescence labeling of individual, living cells which can be imaged using conventional GFP filter sets. Photoactivation can also be efficiently achieved using 2-photon excitation at ~750nm and imaged after photoconversion using excitation at ~950nm. |
| Molecular Note | The R26 PA-GFP::NLS targeting vector was designed with (from 5' to 3') the CMV-IE enhancer/chickenbeta-actin hybrid promoter (CAG; from the pCAGGS vector), a loxP-flanked neomycin-beta-galactosidase fusion protein (betageo) with a STOP codon, a photoactivatable green fluorescent protein (PA-GFP) fused to a nuclear localization signal (NLS), and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability). |
