B6;129P2-Gt(ROSA)26Sor^{tm1(CAG-RABVgp4,-TVA)Arenk}/J

Stock number: 024708
Product name: RphiGT
Strain synonyms: RΦGT

Description

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.

Detailed description

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.

Development

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.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(CAG-RABVgp4,-TVA)Arenk}
Name: targeted mutation 1, Benjamin Arenkiel
MGI accession ID: MGI:5550559
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129P2/OlaHsd
Expressed genes: TVA,subgroup A avian leukosis virus receptor,
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.