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. 2010 Aug 26;17(5):293–301. doi: 10.1093/dnares/dsq020

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© The Author 2010. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Schematic diagram of construction of the tet-O HAC-based vector for regulated expression of genes. (A) The tet-O HAC was constructed in human HT1080 cells. HT1080 carrying the tet-O HAC and mouse A9 cells were fused. (B) Hybrid cells carrying the tet-O HAC were transferred into chicken DT40 cells by MMCT. (C) The loxP/HPRT cassette was inserted into the tet-O HAC by homologous recombination in DT40 cells. (D) The tet-O HAC vector was transferred into CHO cells deficient in HPRT. (E) The EGFP/HPRT cassette was introduced into the HAC by Cre/loxP recombination in CHO cells. (F) The tet-O-EGFP HAC was transferred into human HT1080 cells by MMCT. The tet-O HAC was destabilized by expression of a chromatin modifier gene fused with tet-R.