The invertor knock-in conditional chromosomal translocation mimic

Abstract

Knock-in models of tumor-specific chromosomal translocations can generate lethal mutations. To circumvent this, a new conditional gene fusion model has been developed (invertor mice) and exemplified with the Ews-ERG fusion oncogene. An ERG segment, flanked by loxP sites, was knocked in to an intron of the Ews gene but in an inverted transcription orientation and lineage-specific Ews-ERG fusion created by Cre-mediated inversion. This invertor method is a completely conditional approach, applicable to any gene fusion, to emulate effects of translocations found in human cancers.

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Figure 1: Strategy for generating Ews-ERG invertor gene by homologous recombination.
Figure 2: Leukemia characteristics of Ews-ERG invertor mice.

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Acknowledgements

This work was supported by the Medical Research Council. R.C. was the recipient of a Leukemia Research Fund Gordon Pillar Studentship, M.M. is the recipient of a fellowship from the German Research Foundation, and M.N.L. was the recipient of a Kay Kendall fellowship. We are very grateful to A. McKenzie for the frt-flanked PGK-neo clone. We would also like to thank G. King, A. Middleton and C. Pearce for animal husbandry and A. Lenton for the illustration work.

Author information

Correspondence to Terence H Rabbitts.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Targeting cassettes for invertor or knock-in cDNA cloning. (PDF 184 kb)

Supplementary Fig. 2

Strategy for construction of the invertor cDNA cassette. (PDF 212 kb)

Supplementary Fig. 3

Cassettes for knock-in of the Cre recombinase gene into targeting vectors for gene specific expression. (PDF 190 kb)

Supplementary Fig. 4

ERG cDNA sequence for invertor knock-in targeting clones (PDF 68 kb)

Supplementary Methods (PDF 89 kb)

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