When used in conjunction with a mutant pseudotyped (EnvA) rabies virus reporter, Rabies-SADdeltaG-EGFP, and an appropriate cre-expressing strain, this RΦGT mouse line is useful for monosynaptic neuron tracing.
Benjamin R. Arenkiel, Baylor College of Medicine
These mice contain a recombinant rabies glycoprotein G gene, RABVgp4, and TVA gene (avian leukosis and sarcoma virus subgroup A receptor, chicken) under the control of CAG (CMV enhancer/chicken beta-actin promoter) inserted into the Gt(ROSA)26Sor locus. Expression of RABVgp4 and TVA is blocked by a loxP-flanked STOP fragment placed between the construct sequence and the Gt(ROSA)26Sor promoter.
When used in conjunction with a mutant ASLV-A pseudotyped (EnvA) rabies virus GFP vector, Rabies- SADΔG-EGFP(EnvA)-EGFP, this RΦGT strain serves as a reporter strain for monosynaptic neuron tracing. TVA expression allows the Rabies- SADΔG-EGFP(EnvA)-EGFP to enter the cell, while expression of the rabies virus glycoprotein allows spreading of the viral particles to other cells with direct synaptic contact.
Secondarily infected cells will not express the rabies virus glycoprotein, thus being unable to further spread the Rabies- SADΔG-EGFP(EnvA)-EGFP particle. There is no spreading of the Rabies- SADΔG-EGFP(EnvA)-EGFP without Cre recombinase activity.
Mice that are homozygous for the targeted mutation are viable and fertile. During backcrossing, the Y chromosome was not fixed to the C57BL/6 genetic background.
A targeting vector containing a floxed STOP cassette, recombinant rabies glycoprotein G, IRES sequence and TVA (avian leukosis and sarcoma virus subgroup A receptor, chicken) under the control of CAG (CMV enhancer/chicken beta-actin promoter) was inserted into the Gt(ROSA)26Sor locus. The construct was electroporated into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The donating investigator reports that mice were then backcrossed to C57BL/6 for 5 generations (see SNP note below). During backcrossing, the Y chromosome was not fixed to the C57BL/6 genetic background. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
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. Three of the 27 markers throughout the genome were seregating, suggesting an incomplete backcross.
|Expressed Gene||TVA, subgroup A avian leukosis virus receptor,|
|Site of Expression|
|Allele Name||targeted mutation 1, Benjamin Arenkiel|
|Allele Type||Targeted (Conditional ready (e.g. floxed), Reporter, Inserted expressed sequence)|
|Allele Synonym(s)||targeted mutation 1, Benjamin Arenkiel; Gt(ROSA)26Sortm1(CAG-RABVgp4,-TVA)Arenk|
|Gene Symbol and Name||Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano|
|Gene Synonym(s)||beta geo; ROSA26; R26; Gtrgeo26; Gtrosa26; Gtrgeo26; Gtrosa26; AV258896; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; SETD5-AS1; Thumpd3as1|
|Promoter||CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter,|
|Expressed Gene||TVA, subgroup A avian leukosis virus receptor,|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||The targeting vector contains a loxP-flanked STOP cassette, recombinant rabies glycoprotein G, IRES sequence and TVA (avian leukosis and sarcoma virus subgroup A receptor, chicken) under the control of CAG (CMV enhancer/chicken beta-actin promoter). Expression of the rabies glycoprotein is blocked by the loxP-flanked STOP fragment.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the RphiGT mouse strain in a publication, please cite the originating article(s) and include JAX stock #024708 in your Materials and Methods section.
|Heterozygous for Gt(ROSA)26Sor<tm1(CAG-RABVgp4,-TVA)Arenk>|
We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.
Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.
|Frozen Mouse Embryo||$2,595.00 per straw or vial|
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