The construction of multi-chromatin inducible human artificial chromosome (HAC)

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At the human centromere, various chromatins (e.g. centromere chromatin, heterochromatin) assemble on the highly repetitive DNA (alphoid DNA). Is has been suggested that each chromatin is important for centromere functions, however, the accumulation mechanism of different chromatins on the same DNA and the mechanism of maintenance and regulation of centromere activity are still unknown. From the analyses of a human artificial chromosome (HAC) containing tetracycline operator (tetO), it has been suggested that the balance between various chromatins on centromere directly affected the centromere activity. To analyze the roles of chromatins in detail, we constructed another synthetic alphoid DNA containing lactose operator (lacO). On the tandem array of tetO / lacO centromere DNA, various chromatin modifications could be induced individually by the tethering of tetR- or lacI- fusion proteins.