Kelleher Z T, Fu H, Livanos E, Wendelburg B, Gulino S, Vos J M
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599, USA.
Nat Biotechnol. 1998 Aug;16(8):762-8. doi: 10.1038/nbt0898-762.
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.
我们描述了将100 - 200 kb的自我复制环状人类微型染色体从人类细胞转移到小鼠细胞中的微细胞融合技术。通过将人类疱疹病毒——爱泼斯坦-巴尔病毒的170 kb潜伏双链DNA基因组导入非允许性啮齿动物细胞，阐述了这种实验方法。利用这种种间转移策略，携带95 - 105 kb人类DNA的环状附加体在小鼠A9细胞中以低拷贝数成功建立。选定的附加体稳定维持了6个月，未选定的附加体在每个细胞分裂周期的附加体保留率为95%。小鼠人工附加体染色体系统的建立应有助于在啮齿动物遗传背景下对大型人类DNA进行进化和治疗研究。
