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Molecular and Cellular Biology, May 2004, p. 3794-3803, Vol. 24, No. 9
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.9.3794-3803.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Insertion of the ßGeo Promoter Trap into the Fem1c Gene of ROSA3 Mice

Cassandra L. Schlamp,1 Andrew T. Thliveris,1 Yan Li,1 Louis P. Kohl,1 Claudia Knop,1 Joel A. Dietz,1 Inna V. Larsen,1 Pascal Imesch,1 Lawrence H. Pinto,2 and Robert W. Nickells1*

Department of Ophthalmology and Visual Sciences, University of Wisconsin, Madison, Wisconsin 53704,1 Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 602082

Received 31 October 2003/ Returned for modification 2 December 2003/ Accepted 9 February 2004

ROSA3 mice were developed by retroviral insertion of the ßGeo gene trap vector. Adult ROSA3 mice exhibit widespread expression of the trap gene in epithelial cells found in most organs. In the central nervous system the highest expression of ßGeo is found in CA1 pyramidal cells of the hippocampus, Purkinje cells of the cerebellum, and ganglion cells of the retina. Characterization of the genomic insertion site for ßGeo in ROSA3 mice shows that the trap vector is located in the first intron of Fem1c, a gene homologous to the sex-determining gene fem-1 of Caenorhabditis elegans. Transcription of the Rosa3 allele (R3) yields a spliced message that includes the first exon of Fem1c and the ßGeo coding region. Although normal processing of the Fem1c transcript is disrupted in homozygous Rosa3 (Fem1cR3/R3) mice, some tissues show low levels of a partially processed transcript containing exons 2 and 3. Since the entire coding region of Fem1c is located in these two exons, Fem1cR3/R3 mice may still be able to express a putative FEM1C protein. To this extent, Fem1cR3/R3 mice show no adverse effects in their sexual development or fertility or in the attenuation of neuronal cell death, another function that has been attributed to both fem-1 and a second mouse homolog, Fem1b. Examination of ßGeo expression in ganglion cells after exposure to damaging stimuli indicates that protein levels are rapidly depleted prior to cell death, making the ßGeo reporter gene a potentially useful marker to study early molecular events in damaged neurons.

* Corresponding author. Mailing address: Department of Ophthalmology and Visual Sciences, University of Wisconsin, 6640 MSC, 1300 University Ave., Madison, WI 53706. Phone: (608) 265-6037. Fax: (608) 262-0479. E-mail: nickells{at}wisc.edu.

Molecular and Cellular Biology, May 2004, p. 3794-3803, Vol. 24, No. 9
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.9.3794-3803.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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