Formation of de novo centromeres and construction of first-generation human artificial microchromosomes

Nat Genet. 1997 Apr;15(4):345-55. doi: 10.1038/ng0497-345.


We have combined long synthetic arrays of alpha satellite DNA with telomeric DNA and genomic DNA to generate artificial chromosomes in human HT1080 cells. The resulting linear microchromosomes contain exogenous alpha satellite DNA, are mitotically and cytogenetically stable in the absence of selection for up to six months in culture, bind centromere proteins specific for active centromeres, and are estimated to be 6-10 megabases in size, approximately one-fifth to one-tenth the size of endogenous human chromosomes. We conclude that this strategy results in the formation of de novo centromere activity and that the microchromosomes so generated contain all of the sequence elements required for stable mitotic chromosome segregation and maintenance. This first-generation system for the construction of human artificial chromosomes should be suitable for dissecting the sequence requirements of human centromeres, as well as developing constructs useful for therapeutic applications.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Centromere / genetics
  • Chromosomal Proteins, Non-Histone / analysis
  • Chromosomes, Human / genetics*
  • DNA, Satellite / genetics*
  • Fibrosarcoma
  • Genetic Vectors / genetics*
  • Humans
  • Telomere / genetics
  • Transfection
  • Tumor Cells, Cultured


  • Chromosomal Proteins, Non-Histone
  • DNA, Satellite
  • centromere protein C
  • centromere protein E