Abstract
Complex chromosomal rearrangements (deletions, inversions, translocations) are a hallmark of human tumour cells. Yet, the generation of animal models for gross chromosomal abnormalities still presents a formidable challenge. Here, we describe a versatile procedure for chromosomal engineering that was used to generate an ES cell line with a megabase deletion encompassing the tumour suppressor gene neurofibromatosis-1 (Nf-1) on mouse chromosome 11, which is often deleted in tumours of neural crest origin. Homologous recombination into sites flanking Nf-1 was used to introduce artificial sequences (triple-helix, loxP, vector backbone) that can be employed for in vitro recovery of intervening sequences or the generation of in vivo deletions. This strategy may be developed into a scheme by which large chromosomal regions with precisely defined end points may be excised from mammalian cells and reintroduced after suitable in vitro modification.
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Acknowledgements
We thank Drs M Nehls and M Messerle for the initial isolation and characterization of genomic clones containing markers D11Bhm100 and D11Bhm109, Dr B Sauer for provision of the GFP-cre expression plasmid, and M Sator-Schmitt and H Kohler for excellent technical assistance. This work was initiated at the German Cancer Research Center, Heidelberg, and completed at the Max-Planck-Institute for Immunobiology, Freiburg. Financial support from the Deutsche Forschungsgemeinschaft is gratefully acknowledged.
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Schlake, T., Schupp, I., Kutsche, K. et al. Predetermined chromosomal deletion encompassing the Nf-1 gene. Oncogene 18, 6078–6082 (1999). https://doi.org/10.1038/sj.onc.1203021
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DOI: https://doi.org/10.1038/sj.onc.1203021
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