The new transgenic mouse core: no longer a facility just for making mice
Cornell University, Итак, New York, United StatesLab Animal (Impact Factor: 0.74). 12/2006; 35(10):27-34. DOI: 10.1038/laban1106-27
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.
- [Show abstract] [Hide abstract]
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 · 2.30 Impact Factor
- [Show abstract] [Hide abstract]
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 · 9.11 Impact Factor
- Nature Genetics 02/1999; 21(1):70-1. DOI:10.1038/5007 · 29.35 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.