Epstein-Barr-based episomal chromosomes shuttle 100 kb of self-replicating circular human DNA in mouse cells.
We describe the microcell fusion transfer of 100-200 kb self-replicating circular human minichromosomes from human into mouse cells. This experimental approach is illustrated through the shutting of the latent 170 kb double-stranded DNA genome from the human herpesvirus, Epstein-Barr virus, into nonpermissive rodent cells. Using this interspecies transfer strategy, circular episomes carrying 95-105 kb of human DNA were successfully established at low copy number in mouse A9 cells. Selected episomes were stably maintained for 6 months, and unselected episomes were characterized by a 95% episomal retention per cell division. The establishment of a mouse artificial episomal chromosome system should facilitate evolutionary and therapeutic studies of large human DNA in rodent genetic backgrounds.
Kelleher
Z T
ZT
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599, USA.
Fu
H
H
Livanos
E
E
Wendelburg
B
B
Gulino
S
S
Vos
J M
JM
D000818:Animals; D015139:Blotting, Southern; D002459:Cell Fusion; D002460:Cell Line; D002871:Chromosome Banding; D002877:Chromosomes, Human / Q000235:genetics*; D002934:Cinnamates; D004261:DNA Replication; D004270:DNA, Circular / Q000235:genetics; D018628:Gene Dosage; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction
D002934:Cinnamates; D004270:DNA, Circular; D006921:Hygromycin B; C026273:hygromycin A