These Rap1 double floxed mice possess loxP sites flanking exons in the RAP-related protein (Rap)-1a (Rap1a) and -1b (Rap1b) genes. This strain may be useful for studying synaptic plasticity and changes in cortical inputs to the lateral amygdala.
Alexei Morozov, National Institutes of Health
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Rap1a | RAS-related protein 1a |
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Rap1b | RAS related protein 1b |
These Rap1 double floxed mice possess loxP sites flanking exons 2-3 of the RAP-related protein-1a (Rap1a) gene and loxP sites flanking exon 1 of the RAP-related protein-1b (Rap1b) gene. RAP1 is a small GTPase that has been found in both presynaptic and postsynaptic terminals. It has been implicated in regulation of AMPA receptor (AMPAR) trafficking, long-term potentiation (LTP) of synaptic transmission in the hippocampus, and spatial learning. Mice that are homozygous for these alleles are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have Rap1a exons 2-3 and Rap1b exon 1 deleted in cre-expressing tissues.
For example, when crossed to a strain expressing Cre recombinase in the forebrain, double Rap1 KO mice exhibit impaired fear learning due to alterations in cortical input to the lateral amygdala.
A targeting vector was designed to insert a single loxP site upstream of exon 2, and a loxP-flanked neomycin resistance (neo) cassette downstream of exon 3 of the RAP-related protein-1a (Rap1a) gene. The construct was electroporated into 129S/SvEv-derived MM13 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exons 2-3, intact floxed-neo cassette, or excision of both exons 2-3 and the neo cassette. Correctly targeted ES cells, containing only the floxed-exons 2-3, were injected into blastocysts and resulting chimeric males were bred with C57BL/6J females.
A second targeting vector was designed to insert a single a loxP-flanked neomycin resistance (neo) cassette upstream of exon 1 and a single loxP site downstream of exon 1 of the RAP-related protein-1b (Rap1b) gene. The construct was electroporated into 129S/SvEv-derived MM13 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exon 1, intact floxed-neo cassette, or excision of both exon 1 and the neo cassette. Correctly targeted ES cells, containing only the floxed-exon 1, were injected into blastocysts and resulting chimeric males were bred with C57BL/6J females. Resulting offspring were bred to Rap1a floxed mice, and double floxed mice were maintained on a mixed C57BL/6J;129S background.
Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Allele Name | targeted mutation 1, Alexei Morozov |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Rap1Afloxed |
Gene Symbol and Name | Rap1a, RAS-related protein 1a |
Gene Synonym(s) | |
Strain of Origin | 129S/SvEv |
Chromosome | 3 |
Molecular Note | A loxP site was inserted upstream of exon 2 and a loxP-flanked neomycin selection cassette was inserted downstream of exon 3. Transient expression of cre recombinase removed the neomycin cassette, leaving exons 2 and 3 flanked by loxP sites. Correct targeting was confirmed through Southern blots. |
Allele Name | targeted mutation 1, Alexei Morozov |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | Rap1Bfloxed |
Gene Symbol and Name | Rap1b, RAS related protein 1b |
Gene Synonym(s) | |
Strain of Origin | 129S/SvEv |
Chromosome | 10 |
Molecular Note | A loxP-flanked neomycin selection cassette was inserted upstream of exon 1 with an additional loxP site inserted downstream of the exon. Transient expression of cre recombinase removed the neomycin selection cassette leaving exon 1 flanked by loxP sites. Correct targeted was confirmed through Southern blotting. |
When maintaining a live colony, double homozygous mice may be bred together.
When using the B6;129S-Rap1atm1Morz Rap1btm1Morz/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #021066 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Rap1a<tm1Morz>, Heterozygous for Rap1b<tm1Morz |
Frozen Mouse Embryo | B6;129S-Rap1a<tm1Morz> Rap1b<tm1Morz>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S-Rap1a<tm1Morz> Rap1b<tm1Morz>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S-Rap1a<tm1Morz> Rap1b<tm1Morz>/J | $3373.50 |
Frozen Mouse Embryo | B6;129S-Rap1a<tm1Morz> Rap1b<tm1Morz>/J | $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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