GATA3-eGFP reporter cell line
Conditionally immortal, GATA3eGFP cell line from the embryonic mouse inner ear
Contributors
| Inventor | Institute |
|---|---|
| Matthew C Holley | University of Sheffield |
| Cat. #: | 161125 |
|---|---|
| Unit size: | 1×10^6 cells / vial |
| Research Fields: | Cancer;Developmental biology;Drug development;Genetics |
| Organism: | Mouse |
| Tissue: | Embryonic |
| Model: | Transgeneic |
| Morphology: | Inner ear otocyst at embryonic day E10 |
| Cancer Types In Detail: | Not specific |
| Alternate name: | US/VOT- GATA3eGFP; Univ Sheffield/Ventral Otocyst gata3-enhanced green fluorescent protein; GATA3eGFP; VOT- GATA3eGFP; GATA3eGFP reporter cell line |
|---|---|
| Product description: | This cell line is the only GATA3 reporter line capable of efficiently screening extrinsic factors influencing GATA3 expression in high-throughput systems. The GATA3-eGFP reporter cell line expresses GATA3-eGFP via an enhancer that is expressed in a pattern very closely resembling that of native GATA3 during mouse embryonic development. Fusion of GATA3 enhancer to eGFP is a key feature of this cell line allowing high-throughput screening of drugs that modulate the expression level of gata… |
| CRISPR: | No |
| Conditional: | Yes |
| Conditional description: | Conditionally immortal, see production details |
| Production details: | Animal studies were licensed under the UK Home Office, Animals (Scientific Procedures) Act 1986, and had prior approval of The University of Sheffield Ethical Review Committee. Tissue was harvested from animals culled by a Schedule 1 method. GATA3eGFP mice were a kind gift of Dr Stavros Malos from the Cyprus Institute of Neurology and Genetics and were generated by Bacterial Artificial Chromosome (BAC) recombination [Panayi et al., 2010]. For characterization of embryos, homozygous GATA3… |
| Additional notes: | The expression of eGFP correlates with that of endogenous gata3 in V2c interneurons within the spinal cord from embryonic days E10.5-11.5 [Panayi et al., 2010]. It was found that in freshly dissected or unfixed, cryosectioned tissue it correlated with endogenous expression of gata3 in many other tissues throughout embryonic development. At E10.5 these included the midline dorsal aorta, mesonephric ridge, endothelial cells around the region of the neural lumen, the lens of the eye and specific neurons of the central nervous system. At E12.5 they included the vomeronasal organ and neurons in the mesencephalon, pons and the diencephalon. Images of the whole head clearly revealed expression in the olfactory region, lens, midbrain, hindbrain and spinal cord. A similar pattern was observed at E14.5, including ventral spinal cord, trigeminal ganglion, thyroid glands and tongue. At E16.5, eGFP was observed in the diencephalon, pons, spinal cord, lens, tongue, thymus, carotid arteries, olfactory region and whisker follicles. |
| Model description: | The mice were derived from a cross between two different transgenic mouse lines. One mouse line stably expressed the H-2Kb-tsA58 transgene in which a temperature-sensitive variant of the SV40 large T-antigen was expressed under the control of an inducible promoter driven by gamma-interferon. The second, GATA3eGFP BAC-transgenic mouse line expressed eGFP under the control of the GATA3 promoter. |
| disease: | Development |
| Format: | Frozen |
|---|---|
| Storage conditions: | Liquid Nitrogen |
| Shipping conditions: | Dry ice |
| Growth medium: | Cell culture depends on the experiment and these cells can be cultured under different conditions [eg Milo et al. 2009; Lawoko et al. 2004; Rivolta & Holley, 2002; Lawlor et al., 1999]. Under immortalising conditions they will continue to proliferate so the temperature and level of g-interferon should be managed for optimal growth. The cells can be cultured both in serum and serum free conditions. In serum they can be cultured in MEM (Invitrogen-GIBCO, Paisley, UK) and 10% fetal bov… |
| Mycoplasma free: | Yes |
| Biosafety level: | 1 |
| References: |
Panayi et.al. 2010. J Neurosci. 30(37):12274-80. PMID: 20844123 Milo et.al. 2009. PLoS One. 4(9):e7144. PMID: 19774072 Kurek et. al. 2007. Development. 134(2):261-72. PMID: 17151017 Lawoko-Kerali et. al. 2004. Dev Dyn. 231(4):801-14. PMID: 15499550 Lawoko-Kerali et. al. 2002. J Comp Neurol. 442(4):378-91. PMID: 11793341 Patient and McGhee 2002. Curr Opin Genet Dev. 12(4):416-22. PMID: 12100886 Rivolta and Holley 2002. J Neurobiol. 53(2):306-18. PMID: 12382283 Karis et.al. 2001. J… |
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