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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Abstract

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
Volume98
Issue number10
DOIs
Publication statusPublished - 8 May 2001

Cite this

Saffery, R., Wong, L. H., Irvine, D. V., Bateman, M. A., Griffiths, B., Cutts, S. M., Cancilla, M. R., Cendron, A. C., Stafford, A. J., & Choo, K. H. A. (2001). Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation. Proceedings of the National Academy of Sciences of the United States of America, 98(10), 5705-5710. https://doi.org/10.1073/pnas.091468498