Authors' Affiliations: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford; Leeds Institute of Molecular Medicine, Wellcome Trust Brenner Building, St. James's University Hospital, University of Leeds, Leeds, United Kingdom; and Molecular Cytogenetics Group, Spanish National Cancer Research Center (CNIO), Melchor Fernandez Almagro, Madrid, Spain.
Cancer Res. 2014 Mar 1;74(5):1588-97. doi: 10.1158/0008-5472.CAN-13-1783. Epub 2014 Jan 13.
The discovery of chromosomal translocations in leukemia/lymphoma and sarcomas presaged a widespread discovery in epithelial tumors. With the advent of new-generation whole-genome sequencing, many consistent chromosomal abnormalities have been described together with putative driver and passenger mutations. The multiple genetic changes required in mouse models to assess the interrelationship of abnormalities and other mutations are severe limitations. Here, we show that sequential gene targeting of embryonic stem cells can be used to yield progenitor cells to generate chimeric offspring carrying all the genetic changes needed for cell-specific cancer. Illustrating the technology, we show that MLL-ENL fusion is sufficient for lethal leukocytosis and proof of genome integrity comes from germline transmission of the sequentially targeted alleles. This accelerated technology leads to a reduction in mouse numbers (contributing significantly to the 3Rs), allows fluorescence tagging of cancer-initiating cells, and provides a flexible platform for interrogating the interaction of chromosomal abnormalities with mutations.
染色体易位在白血病/淋巴瘤和肉瘤中的发现预示着上皮性肿瘤的广泛发现。随着新一代全基因组测序的出现，许多一致的染色体异常与推定的驱动和乘客突变一起被描述。在小鼠模型中评估异常和其他突变的相互关系需要多种遗传变化，这是严重的限制。在这里，我们表明，胚胎干细胞的顺序基因靶向可以用于产生祖细胞，以产生携带所有遗传变化的嵌合后代，这些变化是细胞特异性癌症所必需的。为了说明这项技术，我们表明，MLL-ENL 融合足以导致致命性白细胞增多症，而基因组完整性的证明来自顺序靶向等位基因的种系传递。这种加速的技术减少了小鼠数量（对 3R 有重要贡献），允许对起始癌细胞进行荧光标记，并为研究染色体异常与突变的相互作用提供了一个灵活的平台。
