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Chromosomes

Introducing HAC

Nature Reviews Molecular Cell Biology volume 9, page 351 (2008)Cite this article

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Previously, artificial chromosomes that were introduced into human cells contained authentic α-satellite sequences — repetitive DNA that is found at the centromeres of human cells — that contained a DNA-binding site for the centromere protein CENP-B (the CENP-B box). The newly engineered HAC is based on a synthetic repeated DNA sequence called the alphoidtetO dimer. One part of the dimer consists of the natural α-satellite containing the CENP-B box sequences; the other part contains a synthetic α-satellite sequence with the CENP-B box replaced by a tetracycline repressor (tetR) binding site (tetO), which can be used to target desired tetR fusion proteins into the kinetochore.

First, the authors confirmed that the alphoidtetO HAC assembled with kinetochore proteins and segregated correctly as an independent chromosome during mitosis. Fluorescent proteins fused to tetR were efficiently targeted to alphoidtetO and did not affect kinetochore function. Furthermore, chromatin immunoprecipitation (ChIP) analysis showed that key markers of centromeric chromatin, such as the centromere-specific histone variant CENP-A and dimethylated Lys4 on histone H3 (H3K4me2), were present at the alphoidtetO HAC.

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References

ORIGINAL RESEARCH PAPER

  1. Nakano, M. et al. Inactivation of a human kinetochore by specific targeting of chromatin modifiers. Dev. Cell 14, 507–522 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

FURTHER READING

  1. Cheeseman, I. M. & Desai, A. Molecular architecture of the kinetochore–microtubule interface. Nature Rev. Mol. Cell Biol. 9, 33–46 (2008)

    Article  CAS  Google Scholar 

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Authors
  1. Francesca Cesari
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Cesari, F. Introducing HAC. Nat Rev Mol Cell Biol 9, 351 (2008). https://doi.org/10.1038/nrm2392

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  • DOI: https://doi.org/10.1038/nrm2392

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