Ferritin, the iron storage protein, was recently suggested to be a candidate reporter for the detection of gene expression by magnetic resonance imaging (MRI). Here we report the generation of TET:EGFP-HAferritin (tet-hfer) transgenic mice, in which tissue-specific inducible transcriptional regulation of expression of the heavy chain of ferritin could be detected in vivo by MRI. We show organ specificity by mating the tet-hfer mice with transgenic mice expressing tetracycline transactivator (tTA) in liver hepatocytes and in vascular endothelial cells. Tetracycline-regulated overexpression of ferritin resulted in specific alterations of the transverse relaxation rate (R2) of water. Transgene-dependent changes in R2 were detectable by MRI in adult mice, and we also found fetal developmental induction of transgene expression in utero. Thus, the tet-hfer MRI reporter mice provide a new transgenic mouse platform for in vivo molecular imaging of reporter gene expression by MRI during both embryonic and adult life.
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We would like to acknowledge helpful discussions with A. Koretsky, P. Bendel, E. Keshet and Y. Dor. We would like to thank G. Damari and E. Ino for their help in generation and breeding of the transgenic mice. This work was supported by the Israel Science Foundation 391/02 and by the Minerva Foundation (to M.N.).
The authors declare no competing financial interests.
Histological analysis of multiple organs in male VE cadherin h-fer mice. (PDF 136 kb)
Ferritin expression during embryonic development. (PDF 301 kb)
Prussian Blue staining of ferritin iron. (PDF 147 kb)
Contrast enhanced 3D gradient echo MRI of E13.5 pregnant mouse. The dataset was acquired at the end of the spin echo measurement of R2 relaxation, after intravenous administration of biotin-BSA-GdDTPA. Hyperintense areas show the maternal vasculature including maternal circulation in the placenta (same mouse as Figure 3). (AVI 885 kb)
Maximal intensity projection revealing the placenta of E13.5 pregnancy. Movie was derived from the 3D contrast enhanced gradient echo dataset of movie S3. (AVI 652 kb)
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Cohen, B., Ziv, K., Plaks, V. et al. MRI detection of transcriptional regulation of gene expression in transgenic mice. Nat Med 13, 498–503 (2007). https://doi.org/10.1038/nm1497
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