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首页> 外文期刊>Endocrinology >Pituitary Tumors and Immortalized Cell Lines Generated by Cre-Inducible Expression of SV40 T Antigen
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Pituitary Tumors and Immortalized Cell Lines Generated by Cre-Inducible Expression of SV40 T Antigen

机译：通过SV40 T抗原的Cre-Imcuce表达产生垂体肿瘤和永生化细胞系

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摘要

Targeted oncogenesis is the process of driving tumor formation by engineering transgenic mice that express an oncogene under the control of a cell-type specific promoter. Such tumors can be adapted to cell culture, providing immortalized cell lines. To make it feasible to follow the process of tumorigenesis and increase the opportunity for generating cell lines, we developed a mouse strain that expresses SV40 T antigens in response to Cre-recombinase. Using CRISPR/Cas9 we inserted a cassette with coding sequences for SV40T antigens and an internal ribosome entry site with green fluorescent protein cassette (IRES-GFP) into the Rosa26 locus, downstream from a stop sequence flanked by loxP sites: Rosa26(LSL-SV40-GFP) These mice were mated with previously established Prop1-cre and Tshb-cre transgenic lines. Both the Rosa26(LSL-SV40-GFP/+); Prop1-cre and Rosa264(LSL-SV40-GFP/+); Tshb-cre mice developed fully penetrant dwarfism and large tumors by 4 weeks.Tumors from both of these mouse lines were adapted to growth in cell culture. We have established a progenitor-like cell line (PIT-P1) that expresses Sox2 and Pitx1, and a thyrotrope-like cell line (PIT-T1) that expresses Pou1f1 and Cga.These studies demonstrate the utility of the novel, Rosa26(LSL-SV40-GFP) mouse line for reliable targeted oncogenesis and development of unique cell lines.
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