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.
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599, USA.
Z T
ZT
Kelleher
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H
Fu
E
E
Livanos
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B
Wendelburg
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S
Gulino
J M
JM
Vos
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