A photactivatable variant of Aequorea victoria green fluorescent protein was placed under the regulatory control of the human ubiquitin C (UBC) promoter in this transgenic strain. Photoactivation enables rapid, stable fluorescent labeling of living cells with great microanatomical precision.
Michel C Nussenzweig, The Rockefeller University
Genetic Background | Generation |
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?+pN2
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Allele Type | Gene Symbol | Gene Name |
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Not Applicable | Ptprc | protein tyrosine phosphatase, receptor type, C |
Allele Type |
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Transgenic (Reporter) |
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A photactivatable (PA) T203H variant of Aequorea victoria green fluorescent protein (GFP) was placed under the regulatory control of the widely-expressed human ubiquitin C (UBC) promoter in this transgenic strain. Peak excitation wavelength of PA-GFP shifts to ~495 nm upon one-photon irradiation at ~415 nm or two-photon irradiation at ~720-840 nm.
Cells can be photoactivated within intact lymph nodes with great microanatomical precision (~10 microns in the Z dimension, or close to one cell diameter) by two-photon irradiation at 830 nm, and subsequently identified by two-photon excitation at 940 nm or flow cytometry using a conventional 488 nm laser. After a brief recovery period, the migration of photoactivated naive T and germinal center B cells is indistinguishable from that of control cells. The half-life of photoactivated PA-GFP in naive B cells is estimated to be 30 hours, a figure consistent with previous estimates for the half-life of GFP in living cells.
These mice express high levels in all hematopoietic cell populations. Expression in other tissues has not been evaluated. Mutant mice are viable, fertile, and produce normal size litters. Homozygous mice express approximately twice as much protein as hemizygotes, as determined by flow cytometry. Photoactivation enables rapid, stable fluorescent labeling of living cells with great microanatomical precision.
A transgenic vector carrying photoactivatable green fluorescent protein (PA-GFP, T203H variant) under the transcriptional control of the human ubiquitin C promoter was injected into C57BL/6 embryos. Resultant mice were crossed with a congenic strain on a C57BL/6 background to introduce a Ptprca (CD45.1/Ly5.1) allele (see Stock No. 002014). These mice were maintained on a C57BL/6 genetic background by the donating laboratory (see SNP note below).
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | Fluorescent labeling is expressed in high levels in all hematopoietic cell populations. |
Allele Name | a variant |
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Allele Type | Not Applicable |
Allele Synonym(s) | CD45.1; Ly5a; PtprcSJL |
Gene Symbol and Name | Ptprc, protein tyrosine phosphatase, receptor type, C |
Gene Synonym(s) | |
Site of Expression | Widely expressed on all adaptive and innate immune cells. |
Strain of Origin | Not Applicable |
Chromosome | 1 |
Molecular Note | Ptprca is found in strains SJL/J, STS/A, and DA. Ptprcb is found in strains C57BL/6, C3H/An, DBA/2, AKR, and many others (J:13367, J:12054, J:12077, J:8603). Twelve nucleotide differences between the a and b alleles have been identified. These base substitutions correspond to five amino-acid changes within the extracellular domain of the encoded protein. These amino-acid differences are clustered in a region that also contains the greatest divergence between mouse and rat sequences (J:22485). Note that the allele designations originally described were reversed in 1987 (J:8603); all publications prior to 1987 show SJL/J, STS/A, and DA as having the b allele and the C57BL/6J group as having the a allele (J:22341). |
Allele Name | transgene insertion 1, Michel C Nussenzweig |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | |
Gene Symbol and Name | Tg(UBC-PA-GFP)1Mnz, transgene insertion 1, Michel C Nussenzweig |
Gene Synonym(s) | |
Promoter | UBC, ubiquitin C, human |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | Fluorescent labeling is expressed in high levels in all hematopoietic cell populations. |
Strain of Origin | C57BL/6J |
Chromosome | UN |
Molecular Note | This transgene comprises 1.2 kb of 5'-flanking DNA (nucleotides -1225 to -6) of the human ubiquitin C gene, which is expressed in a broad range of cell types, including all hematopoietic cells, driving expression of a photoactivatable green fluorescent protein cDNA, which is followed by both the SV40 and the bovine growth hormone polyadenylation signals. PA-GFP differs from EGFP by the substitution of histidine for threonine at amino acid position 203 (T203H); this modification results in ~100-fold enhanced fluorescence of PA-GFP upon excitation at 488 nm after preliminary photoactivation with intense 413-nm light, greatly increasing the optical contrast. |
Mice homozygous or hemizygous for the transgene are viable and fertile. The donating laboratory genotypes animals using flow cytometry (AmCyan channel in common BD Biosciences LSR cytometers) or through common GFP PCR.
When using the UBC PA-GFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #022486 in your Materials and Methods section.
Service/Product | Description | Price |
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Hemizygous for Tg(UBC-PA-GFP)1Mnz |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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