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

doi:10.1073/pnas.091468498

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

Research output: Contribution to journal › Article › Research › peer-review

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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 language	English
Pages (from-to)	5705-5710
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	98
Issue number	10
DOIs	https://doi.org/10.1073/pnas.091468498
Publication status	Published - 8 May 2001

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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

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title = "Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation",

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.",

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

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Saffery, R, Wong, LH, Irvine, DV, Bateman, MA, Griffiths, B, Cutts, SM, Cancilla, MR, Cendron, AC, Stafford, AJ & Choo, KHA 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, vol. 98, no. 10, pp. 5705-5710. https://doi.org/10.1073/pnas.091468498

Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation. / Saffery, Richard; Wong, Lee H.; Irvine, Danielle V. et al.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 10, 08.05.2001, p. 5705-5710.

Research output: Contribution to journal › Article › Research › peer-review

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AU - Saffery, Richard

AU - Wong, Lee H.

AU - Irvine, Danielle V.

AU - Bateman, Melissa A.

AU - Griffiths, Belinda

AU - Cutts, Suzanne M.

AU - Cancilla, Michael R.

AU - Cendron, Angela C.

AU - Stafford, Angela J.

AU - Choo, K. H Andy

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AB - 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.

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

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