Nature Genetics
8, 33 - 41 (1994)
doi:10.1038/ng0994-33
Human artificial episomal chromosomes for cloning large DNA fragments in human cellsTian-Qiang Sun1, David A. Fenstermacher2
& Jean-Michel H. Vos1, 2, 3, 4
1Department of Biochemistry and Biophysics, CB#;7295, 349 LCCC, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA
2Curriculum in Genetics and Molecular Biology, CB#7295, 349 LCCC, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA
3UNC Lineberger Comprehensive Cancer Center, CB#7295, 349 LCCC, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA
4Correspondence should be addressed to J.-M.H.V. We have developed a human artificial episomal chromosome (HAEC) system, based on the latent replication origin of the large herpes Epstein-Barr virus, for the propagation and stable maintenance of DNA as circular minichromosomes in human cells. Individual HAECs carried human genomic inserts ranging from 60−330 kb and appeared genetically stable. An HAEC library of 1,500 independent clones carrying random human genomic fragments with average sizes of 150−200 kb was established and allowed recovery of the HAEC DNA. Our autologous HAEC system, with human DNA cloned directly in human cells, provides an important tool for functional study of large mammalian DNA regions and gene therapy.
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