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
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|
|Site of Expression|
|Allele Name||transgene insertion 3, Elizabeth M Simpson|
|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|
|Promoter||Pgk1, phosphoglycerate kinase 1, mouse, laboratory|
|Expressed Gene||neo, neomycin resistance, bacterial|
|Strain of Origin||C57BL/6J|
|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.
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.