The new transgenic mouse core: no longer a facility just for making mice
ABSTRACT The times they are a-changin' for GEM and the facilities that maintain them. Core facilities are expanding beyond their original conception as producers of transgenic mice to encompass a wide range of services, including research animal maintenance. In this paper, the authors describe the logistics and administration of the newly dubbed Mouse Genetics Core Facility at the Memorial Sloan-Kettering Cancer Center as a blueprint for other institutions seeking to expand and update their own transgenic cores for research in the twenty-first century.
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ABSTRACT: The Cre/loxPsite-specific recombination system derived from bacteriophage P1 provides a convenient tool for directed modifications of genomes in various organisms. To exploit Cre-mediated manipulation of mouse genomic sequences at the zygote stage, we have developed a transgenic mouse line carrying the CAG-cretransgene in which thecregene is under control of the cytomegalovirus immediate early enhancer-chicken β-actin hybrid (CAG) promoter. The activity of the Cre recombinase at early stages of development was examined by crossing the CAG-cretransgenic mice to another transgenic mouse line carrying a reporter gene construct, CAG-CAT-Z, which directs expression of theE. coli lacZgene upon Cre-mediated excision of theloxP-flanked chloramphenicol acetyltransferase (CAT) gene located between the CAG promoter and thelacZgene. PCR-based analysis of F1 progeny from CAG-cremales × CAG-CAT-Z females showed that transmission of the CAG-cretransgene was accompanied by the complete deletion of the CAT gene of the CAG-CAT-Z transgene in all tissues, and that this deletion was never observed in the progeny without transmission of the CAG-cregene. On the other hand, analysis of F1 mice from CAG-CAT-Z males × CAG-crefemales showed that the CAG-CAT-Z transgene had undergone complete deletion of the CAT gene in all tissues irrespective of the cotransmission of the CAG-cregene. This Cre-mediated recombination in F1 mice occurred before the two-cell stage of embryonic development, as shown by X-gal staining. The results suggest that the CAG-cretransgene is expressed in developing oocytes of CAG-cretransgenic mice, and Cre mRNA and/or protein are retained in mature oocytes irrespective of the transmission of the CAG-cretransgene, resulting in efficient Cre-mediated recombination of paternally derived target genes upon fertilization. The CAG-cretransgenic mouse should serve as a useful tool to introduce prescribed genetic modifications into the mouse embryo at the zygote stage.Biochemical and Biophysical Research Communications 09/1997; 237(2):318-24. DOI:10.1006/bbrc.1997.7111 · 2.28 Impact Factor
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ABSTRACT: Murine embryonic stem (ES) cells are commonly cultured on feeder layers of primary murine embryonic fibroblasts (MEFs). Because gene targeting experiments often involve sequential selection for multiple-drug resistance in single ES cell lines, we have developed a new mouse strain which represents an economical donor for the production of multiple-drug resistant MEFs. MEFs prepared from the DR-4 mouse strain displayed resistance to concentrations of the drugs G418, 6-thioguanine, puromycin and hygromycin well above those used normally for the selection of drug-resistant ES cells.Nucleic Acids Research 10/1997; 25(18):3745-6. DOI:10.1093/nar/25.18.3745 · 8.81 Impact Factor
Nature Genetics 02/1999; 21(1):70-1. DOI:10.1038/5007 · 29.65 Impact Factor