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Nucleic Acids Res. 2011 Sep 1;39(16):e106. doi: 10.1093/nar/gkr442. Epub 2011 Jun 7.

Gene amplification system based on double rolling-circle replication as a model for oncogene-type amplification.

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National Institute for Basic Biology, Department of Basic Biology, The Graduate University for Advanced Studies (Sokendai), Myodaiji, Okazaki, Aichi, 444-8585, Japan.


Gene amplification contributes to a variety of biological phenomena, including malignant progression and drug resistance. However, details of the molecular mechanisms remain to be determined. Here, we have developed a gene amplification system in yeast and mammalian cells that is based on double rolling-circle replication (DRCR). Cre-lox system is used to efficiently induce DRCR utilizing a recombinational process coupled with replication. This system shows distinctive features seen in amplification of oncogenes and drug-resistance genes: (i) intra- and extrachromosomal amplification, (ii) intensive chromosome rearrangement and (iii) scattered-type amplification resembling those seen in cancer cells. This system can serve as a model for amplification of oncogenes and drug-resistance genes, and improve amplification systems used for making pharmaceutical proteins in mammalian cells.

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