Insertion of the beta Geo promoter trap into the Fem1c gene of ROSA3 mice.
ROSA3 mice were developed by retroviral insertion of the beta Geo 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 beta Geo 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 beta Geo 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 beta Geo coding region. Although normal processing of the Fem1c transcript is disrupted in homozygous Rosa3 (Fem1c(R3/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, Fem1c(R3/R3) mice may still be able to express a putative FEM1C protein. To this extent, Fem1c(R3/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 beta Geo expression in ganglion cells after exposure to damaging stimuli indicates that protein levels are rapidly depleted prior to cell death, making the beta Geo reporter gene a potentially useful marker to study early molecular events in damaged neurons.
Schlamp
Cassandra L
CL
Department of Ophthalmology and Visual Sciences, University of Wisconsin, Madison, Wisconsin 53704, USA.
Thliveris
Andrew T
AT
Li
Yan
Y
Kohl
Louis P
LP
Knop
Claudia
C
Dietz
Joel A
JA
Larsen
Inna V
IV
Imesch
Pascal
P
Pinto
Lawrence H
LH
Nickells
Robert W
RW
D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D011401:Promoter Regions, Genetic; D012160:Retina / Q000166:cytology / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D043743:Ubiquitin-Protein Ligase Complexes
D004268:DNA-Binding Proteins; D014157:Transcription Factors; C486646:Fem1c protein, mouse; D043743:Ubiquitin-Protein Ligase Complexes