title: A high G418-resistant neo(R) transgenic mouse and mouse embryonic fibroblast (MEF) feeder layers for cytotoxicity and gene targeting in vivo and in vitro.
abstract: Aminoglycoside antibiotics have been in use since 1944 with the discovery of streptomycin. The aim of this study was to derive a new, highly resistant multicopy neo(R) transgenic mouse strain, named TgN3Ems, by random insertion of the plasmid, pPGKneobpA, and compare the level of drug resistance of wild-type and transgenic mice in vivo and corresponding primary mouse embryonic fibroblasts (MEFs) in vitro to a model neomycin analog, G418. The expression neoR in transgenic animals caused a 5-fold increase in the approximate lethal dose of G418, compared to wild type. No adverse pathological changes were found for the transgenic mice treated with G418, as they all died within minutes after injection. In contrast, the G418 treatment of wild-type mice resulted in a marked liver and kidney toxicity detected microscopically and via increases of serum biomarkers for liver and kidney damage. In addition, there was a mild bone marrow and lymphoid depletion. In in vitro studies, the transgenic MEFs survived 20-fold higher G418 levels, compared to the wild-type MEF cells. Therefore, TgN3Ems transgenic mice could be used as a source of G418-resistant feeder cells for gene targeting. Since the expression of drug-resistance genes in transgenic animals confers resistance to toxicity, the TgN3Ems mice might serve as a tool applicable in drug design.
authors: Harvard School of Public Health, Boston, Massachusetts, USA.
authors: Jiri
authors: J
authors: Aubrecht
authors: Mary E P
authors: ME
authors: Goad
authors: Agnieszka K
authors: AK
authors: Czopik
authors: Charles P
authors: CP
authors: Lerner
authors: Kevin A
authors: KA
authors: Johnson
authors: Elizabeth M
authors: EM
authors: Simpson
authors: Robert H
authors: RH
authors: Schiestl
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D004351:Drug Resistance / Q000235:genetics*; D061252:Feeder Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D018390:Gene Targeting; D005839:Gentamicins / Q000494:pharmacology / Q000633:toxicity*; D019868:Kanamycin Kinase / Q000235:genetics*; D007928:Lethal Dose 50; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010957:Plasmids; D011401:Promoter Regions, Genetic; D023382:Toxicity Tests, Acute; D019076:Transgenes
chemical_list: D005839:Gentamicins; C010680:antibiotic G 418; D019868:Kanamycin Kinase