Novel generation of human satellite DNA-based artificial chromosomes in mammalian cells

E. Csonka, I. Cserpan, K. Fodor, G. Hollo, R. Katona, J. Kereso, T. Praznovszky, B. Szakal, A. Telenius, G. deJong, A. Udvardy, G. Hadlaczky

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Abstract

An in vivo approach has been developed for generation of artificial chromosomes, based on the induction of intrinsic, large-scale amplification mechanisms of mammalian cells. Here, we describe the successful generation of prototype human satellite DNA-based artificial chromosomes via amplification-dependent de novo chromosome formations induced by integration of exogenous DNA sequences into the centromeric/rDNA regions of human acrocentric chromosomes. Subclones with mitotically stable de novo chromosomes were established, which allowed the initial characterization and purification of these artificial chromosomes. Because of the low complexity of their DNA content, they may serve as a useful tool to study the structure and function of higher eukaryotic chromosomes. Human satellite DNA-based artificial chromosomes containing amplified satellite DNA, rDNA, and exogenous DNA sequences were heterochromatic, however, they provided a suitable chromosomal environment for the expression of the integrated exogenous genetic material. We demonstrate that induced de novo chromosome formation is a reproducible and effective methodology in generating artificial chromosomes from predictable sequences of different mammalian species. Satellite DNA-based artificial chromosomes formed by induced large-scale amplifications on the short arm of human acrocentric chromosomes may become safe or low risk vectors in gene therapy.

Original languageEnglish
Pages (from-to)3207-3216
Number of pages10
JournalJournal of cell science
Volume113
Issue number18
Publication statusPublished - Jan 1 2000

Keywords

  • Human satellite DNA-based artificial chromosome
  • Large-scale amplification
  • RDNA

ASJC Scopus subject areas

  • Cell Biology

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    Csonka, E., Cserpan, I., Fodor, K., Hollo, G., Katona, R., Kereso, J., Praznovszky, T., Szakal, B., Telenius, A., deJong, G., Udvardy, A., & Hadlaczky, G. (2000). Novel generation of human satellite DNA-based artificial chromosomes in mammalian cells. Journal of cell science, 113(18), 3207-3216.