Srsf2P95H floxed mice possess loxP sites flanking the coding region of the Srsf2 gene, and also contains a downstream mutant coding region. Removal of the floxed wild type exons allows expression of the mutant sequence. This strain may be useful for studying myelodysplastic syndromes.
Omar Abdel-Wahab, Memorial Sloan Kettering Cancer Center
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed)) | Srsf2 | serine/arginine-rich splicing factor 2 |
Srsf2P95H floxed mice possess loxP sites flanking the endogenous coding region of the serine/arginine-rich splicing factor 2 (Srsf2) gene. They also contain a duplication of the coding region containing a proline to histidine mutation downstream of the 3’ loxP site. SRSF2 encodes a spliceosomal gene associated with the regulation of consititive splicing. Mutations in SRSF2 have been associated with the onset and pathogenesis of myelodysplastic syndromes (MDS) and chronic myelomonocytic leukemia (CMML). Srsf2P95H floxed mice that are heterozygous for this allele are viable and fertile, the donating investigator has not attempted to make homozygotes. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have endogenous exons deleted, and will instead express the mutant Srsf2P95H gene in the cre-expressing tissues.
When bred to B6.Cg-Tg(Mx1-cre)1Cgn/J mice (from Stock No. 003556), expressing Cre recombinase in hematopoietic stem cells (HSCs) after administration of poly(IC), mice develop a mild anemia with macrocytosis and leukopenia, as well as morpholgic dysplasia as seen in human myelodysplastic syndromes.
A targeting vector was designed by Dr. Omar Abdel-Wahab (Memorial Sloan Kettering Cancer Center) to insert a loxP site upstream of exon 1 of the serine/arginine-rich splicing factor 2 (Srsf2) gene. cDNA encoding the sequence of exon 3, and an SV40 polyadenylation sequence bound to a frt-flanked neomycin resistance (neo) cassette, followed by a second loxP site were inserted downstream of exon 2. Duplicate exons 1-2 were inserted downstream of the neo cassette. This second exon 1 contains a point mutation in amino acid 95 which alters the nucleotide sequence from CCG to CAC, resulting in an amino acid change from Proline to Histidine. The cDNA for exon 3 was fused in frame to endogenous exon 2. The construct was electroporated into C57BL/6NTac-derived IC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into Balb/c blastocysts and resulting chimeric mice were bred with B6.Cg-Tg(ACTFLPe)9205Dym/J transgenic mice (Stock No. 005703) to delete the neo cassette. Resulting progeny were crossed to remove the Flp-expressing transgene, and Srsf2P95H floxed offspring were backcrossed to C57BL/6 mice for at least 6 generations (see SNP note below). Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
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, 4 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.
Allele Name | targeted mutation 1.1, Omar Abdel-Wahab |
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Allele Type | Targeted (Conditional ready (e.g. floxed)) |
Allele Synonym(s) | Srsf2P95H |
Gene Symbol and Name | Srsf2, serine/arginine-rich splicing factor 2 |
Gene Synonym(s) | |
Strain of Origin | C57BL/6NTac |
Chromosome | 11 |
Molecular Note | A targeting vector was designed to insert a loxP site upstream of exon 1, cDNA encoding the sequence of exon 3 fused in frame to the endogenous exon 2, followed by an SV40 polyadenylation sequence bound to an FRT-flanked neomycin resistance (neo) cassette and a second loxP site. Duplicate exons 1-2 were inserted downstream of the second loxP site and upstream of the endogenous exon 3. The second exon 1 contains a mutation in amino acid 95 which alters the nucleotide sequence from CCG to CAC, resulting in an amino acid change from Proline to Histidine. Flp-mediated recombination removed the FRT-flanked neo cassette. |
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony, or to C57BL/6J inbred mice (Stock No. 000664). Homozygous mice die prior to weaning.
When using the Srsf2P95H floxed mouse strain in a publication, please cite the originating article(s) and include JAX stock #028376 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Srsf2<tm1.1Oaw> |
Frozen Mouse Embryo | B6J.B6NTac(SJL)-Srsf2<tm1.1Oaw>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6J.B6NTac(SJL)-Srsf2<tm1.1Oaw>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6J.B6NTac(SJL)-Srsf2<tm1.1Oaw>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6J.B6NTac(SJL)-Srsf2<tm1.1Oaw>/J Frozen Embryo | $3373.50 |
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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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