Abstract
Chromosomal deletions, especially nested deletions, are major genetic tools in diploid organisms that facilitate the functional analysis of large chromosomal regions and allow the rapid localization of mutations to specific genetic intervals. In mice, well-characterized overlapping deletions are only available at a few chromosomal loci1,2, partly due to drawbacks of existing methods. Here we exploit the random integration of a retrovirus to generate high-resolution sets of nested deletions around defined loci in embryonic stem (ES) cells, with sizes extending from a few kilobases to several megabases. This approach expands the application of Cre-loxP–based chromosome engineering3 because it not only allows the construction of hundreds of overlapping deletions, but also provides molecular entry points to regions based on the retroviral tags. Our approach can be extended to any region of the mouse genome.
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Acknowledgements
We thank N. Copeland and N. Jenkins for CAST/Ei mice; P. Liu for Hsd17b1 targeting vectors; W. Cai and J. Li for advice on BAC screening; Y. Cheah for generating chimaeras; S. Perez for preparation of the manuscript; and G. Luo, B. Zheng, P. Biggs, D. Thompson and M. Wentland for critical reading of the manuscript. This work was supported by grants from The National Cancer Institute. X.W. is an Associate and A.B. an Investigator with the Howard Hughes Medical Institute.
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Su, H., Wang, X. & Bradley, A. Nested chromosomal deletions induced with retroviral vectors in mice . Nat Genet 24, 92–95 (2000). https://doi.org/10.1038/71756
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DOI: https://doi.org/10.1038/71756
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