Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation

Richard Saffery, Lee H. Wong, Danielle V. Irvine, Melissa A. Bateman, Belinda Griffiths, Suzanne M. Cutts, Michael R. Cancilla, Angela C. Cendron, Angela J. Stafford, K. H Andy Choo

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Neocentromeres (NCs) are fully functional centromeres that arise ectopically in noncentromeric regions lacking α-satellite DNA. Using telomere-associated chromosome truncation, we have produced a series of minichromosomes (MiCs) from a mardel(10) marker chromosome containing a previously characterized human NC. These MiCs range in size from ≅0.7 to 1.8 Mb and contain single-copy intact genomic DNA from the 10q25 region. Two of these NC-based Mi-Cs (NC-MiCs) appear circular whereas one is linear. All demonstrate stability in both structure and mitotic transmission in the absence of drug selection. Presence of a functional NC is shown by binding a host of key centromere-associated proteins. These NC-MiCs provide direct evidence for mitotic segregation function of the NC DNA and represent examples of stable mammalian MiCs lacking centromeric repeats.
Original languageEnglish
Pages (from-to)5705-5710
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number10
Publication statusPublished - 8 May 2001

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