An EGFP sequence knocked into exon 2 of Slc17a8 abolishes gene function in these mice. Useful applications include studying the role of glutamate transporters in the nervous and auditory systems.
Robert Edwards, UCSF
An enhanced green fluorescent protein (EGFP) replaces most of exon 2 of the solute carrier family 17, member 8 (Slc17a8 or Vglut3) gene in these mice. Gene function is abolished. EGFP signal is not detected. Homozygotes are viable, fertile, and normal in size. VGLUT3 is a vesicular glutamate transporter that mediates synaptic transmission by transporting the neurotransmitter, glutamate, into secretory vesicles. In wildtype mice, VGLUT3 is expressed by inner hair cells of the cochlea, by serotonergic neurons, cholinergic neurons, a particular subset of primary sensory afferents (low-threshold mechanoreceptors), a subset of GABAergic interneurons, and a variety of other cell populations, some not known to use glutamate as a transmitter. These VGLUT3 deficient mice are deaf and exhibit epileptic seizures with little change in motor behavior.
A targeting vector was designed to replace most of exon 2 encoding the solute carrier family 17, member 8 (Slc17a8 or Vglut3) gene with an enhanced green fluorescent protein (EGFP) followed by a polyadenylation signal and a floxed-neomycin resistance (neo) cassette. The construct was electroporated into 129P2/Ola-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to C57BL/6 females. Heterozygous offspring were subsequently bred to mice expressing ACTB-cre to delete the neo cassette. The donating investigator reported that these mice were backcrossed at least 10 generations to C57BL/6 background (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation 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. 6 of the 27 markers throughout the genome were segregating between B6J and 129, suggesting an incomplete backcross.
|Allele Name||targeted mutation 1, Robert H Edwards|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Slc17a8, solute carrier family 17 (sodium-dependent inorganic phosphate cotransporter), member 8|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||Most of exon 2 was replaced with an EGFP cassette, with a stop codon and poly-adenylation signal, and floxed neo cassette. Germ-line, cre-mediated recombination was used to remove the neo cassette. The absence of protein product was confirmed by western blot analysis on brain extracts. However, no EGFP signal was detected.|
|Mutations Made By|| |
Robert Edwards, UCSF
When maintaining a live colony, homozygous mice may be bred together.
When using the VGLUT3- mouse strain in a publication, please cite the originating article(s) and include JAX stock #016931 in your Materials and Methods section.