These TgN3Ems transgenic mice express a neomycin resistance cassette (neoR), rendering the mice resistant to the lethal effects of the neomycin analog, G418. These TgN3Ems mice may be useful in drug design.
Elizabeth M Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia
Genetic Background | Generation |
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Allele Type |
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Transgenic (Inserted expressed sequence) |
Homozygous neoR transgenic mice (TgN3Ems) are viable and fertile with no apparent gross phenotype. The pPGKneobpA transgene has the mouse Pgk1 promoter directing widespread (but not necessarily uniform) expression of neoR: this renders TgN3Ems mice highly G418-resistant and results in a 5-fold increase in the approximate lethal dose of G418 (~480 mg/kg) compared to wildtype mice (~103 mg/kg). Treatment with G418 lethal dose (or higher) in transgenic mice leads to rapid onset of decreased movement, respiratory arrest, and death within minutes. Compared to wildtype mouse embryonic fibroblasts (MEFs), TgN3Ems MEFs exhibit greatly diminished G418 sensitivity: wildtype MEFs show essentially no resistance to G418 while TgN3Ems MEFs survive approximately 20-fold higher G418 levels. Importantly, TgN3Ems heterozygotes and TgN3Ems homozygotes are equally resistant to G418. These TgN3Ems transgenic mice may be useful as source of G418-resistant feeder cells for gene targeting; specifically TgN3Ems MEFs may be ideal for two-allele, as well as for standard, targeting due to their high resistance to selection drugs. Additionally, because the expression of drug-resistance genes in transgenic animals confers resistance to toxicity, the TgN3Ems mice may also serve as a tool strain for testing drug design.
This neoR transgenic mouse strain (TgN3Ems) was created by the laboratory of Dr. Elizabeth M. Simpson while at The Jackson Laboratory. The 1.6 kb pPGKneobpA transgene was designed with (from 5' to 3') the mouse 3-phosphoglycerate kinase (Pgk1) promoter, a neomycin resistance cassette (neoR), and a bovine growth hormone polyadenylation site (bpA). The transgene was microinjected into C57BL/6J wildtype mice embryos. The resulting transgenic founder mouse was bred with C57BL/6J mice to establish line 3. TgN3Ems transgenic mice were sent to The Jackson Laboratory Repository in 1995, and have been maintained by breeding with C57BL/6J wildtype mice and/or breeding together as homozygotes for several generations.
Expressed Gene | neo, neomycin resistance, bacterial |
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Site of Expression |
Allele Name | transgene insertion 3, Elizabeth M Simpson |
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Allele Type | Transgenic (Inserted expressed sequence) |
Allele Synonym(s) | neor; PGK neo; TgN3Ems |
Gene Symbol and Name | Tg(pPGKneobpA)3Ems, transgene insertion 3, Elizabeth M Simpson |
Gene Synonym(s) | |
Promoter | Pgk1, phosphoglycerate kinase 1, mouse, laboratory |
Expressed Gene | neo, neomycin resistance, bacterial |
Strain of Origin | C57BL/6J |
Chromosome | UN |
General Note | Heterozygous and homozygous transgenic mice carry G418-resistant cells. |
Molecular Note | The transgene contains a phosphoglycerate kinase 1 promoter and a neomycin resistance cassette. |
Mutations Made By | Elizabeth Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia |
When maintaining a live colony, homozygous mice may be bred together. Alternatively, hemizygous mice may be bred to wildtype siblings or to C57BL/6J inbred mice (Stock No. 000664). The expected coat color from these breedings is Black.
When using the TgN3Ems mouse strain in a publication, please cite the originating article(s) and include JAX stock #002356 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Homozygous for Tg(pPGKneobpA)3Ems, 1 pair minimum |
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