Construction of neocentromere-based human minichromosomes by telomere-associated chromosomal truncation.
Neocentromeres (NCs) are fully functional centromeres that arise ectopically in noncentromeric regions lacking alpha-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 approximately 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.
Saffery
R
R
The Murdoch Children's Research Institute, Royal Children's Hospital, Flemington Road, Melbourne 3052, Australia.
Wong
L H
LH
Irvine
D V
DV
Bateman
M A
MA
Griffiths
B
B
Cutts
S M
SM
Cancilla
M R
MR
Cendron
A C
AC
Stafford
A J
AJ
Choo
K H
KH
D002460:Cell Line; D002503:Centromere; D002877:Chromosomes, Human; D018014:Gene Transfer Techniques; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016615:Telomere