Site-specific integration of adeno-associated virus involves partial duplication of the target locus.

E. Henckaerts, N. Dutheil, N. Zeltner, S. Kattman, E. Kohlbrenner, P. Ward, N. Clement, P. Rebollo, M. Kennedy, G. M. Keller, R. M. Linden
Proceedings of the National Academy of Sciences. 2009-04-16; 106(18): 7571-7576
DOI: 10.1073/pnas.0806821106

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1. Proc Natl Acad Sci U S A. 2009 May 5;106(18):7571-6. doi:
10.1073/pnas.0806821106. Epub 2009 Apr 16.

Site-specific integration of adeno-associated virus involves partial duplication
of the target locus.

Henckaerts E(1), Dutheil N, Zeltner N, Kattman S, Kohlbrenner E, Ward P, Clément
N, Rebollo P, Kennedy M, Keller GM, Linden RM.

Author information:
(1)Department of Infectious Diseases, King’s College London School of Medicine,
London SE1 9RT, United Kingdom.

A variety of viruses establish latency by integrating their genome into the host
genome. The integration event generally occurs in a nonspecific manner,
precluding the prediction of functional consequences from resulting disruptions
of affected host genes. The nonpathogenic adeno-associated virus (AAV) is unique
in its ability to stably integrate in a site-specific manner into the human MBS85
gene. To gain a better understanding of the integration mechanism and the
consequences of MBS85 disruption, we analyzed the molecular structure of AAV
integrants in various latently infected human cell lines. Our study led to the
observation that AAV integration causes an extensive but partial duplication of
the target gene. Intriguingly, the molecular organization of the integrant leaves
the possibility that a functional copy of the disrupted target gene could
potentially be preserved despite the resulting rearrangements. A latently
infected, Mbs85-targeted mouse ES cell line was generated to study the functional
consequences of the observed duplication-based integration mechanism.
AAV-modified ES cell lines continued to self-renew, maintained their multilineage
differentiation potential and contributed successfully to mouse development when
injected into blastocysts. Thus, our study reveals a viral strategy for targeted
genome addition with the apparent absence of functional consequences.

DOI: 10.1073/pnas.0806821106
PMCID: PMC2669786
PMID: 19372372 [Indexed for MEDLINE]

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