Therapeutic gene delivery in human B-lymphoblastoid cells by engineered non-transforming infectious Epstein-Barr virus.

Abstract

The B-lymphotrophic human herpes Epstein-Barr virus (EBV) is a 160-kilobase double-stranded DNA episomal virus carried in a persistent asymptomatic state by more than 90% of the worldwide adult population. We engineered a helper-dependent mini-EBV, with the minimal cis-EBV elements for episomal replication, viral amplification and packaging, for use as a gene delivery system. The therapeutic potential of this system was established by stably transducing B-lymphoblastoid cells from a Fanconi anaemia group C (FA-C) patient with a mini-EBV constitutively expressing the normal FACC cDNA and showing in vitro correction of the FA phenotype. In the absence of selective pressure, episomal expression persisted with a half-life of 30 days in actively growing transduced cells, indicating a retention rate of 98% expression per cell doubling. This work demonstrates the generation of an infectious non-transforming viral vector that can potentially deliver large therapeutic genes efficiently and selectively into human B cells.

• Authors: S Banerjee, E Livanos, J M Vos
• Journal: Nature medicine (Nat Med) Vol. 1 Issue 12 Pg. 1303-8 (Dec 1995) ISSN: 1078-8956 [Print] United States
• PMID: 7489413 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA Primers

Topics

• B-Lymphocytes (cytology)
• Base Sequence
• Cell Line
• Chromosomes (metabolism)
• DNA Primers
• Fanconi Anemia (pathology)
• Genetic Engineering
• Genetic Therapy (methods)
• Genetic Vectors
• Herpesvirus 4, Human (genetics)
• Humans
• Molecular Sequence Data
• Plasmids
• Transduction, Genetic