Ribosomal DNA integrating rAAV-rDNA vectors allow for stable transgene expression

Leszek Lisowski, Ashley Lau, Zhongya Wang, Yue Zhang, Feijie Zhang, Markus Grompe, Mark A. Kay

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

Although recombinant adeno-associated virus (rAAV) vectors are proving to be efficacious in clinical trials, the episomal character of the delivered transgene restricts their effectiveness to use in quiescent tissues, and may not provide lifelong expression. In contrast, integrating vectors enhance the risk of insertional mutagenesis. In an attempt to overcome both of these limitations, we created new rAAV-rDNA vectors, with an expression cassette flanked by ribosomal DNA (rDNA) sequences capable of homologous recombination into genomic rDNA. We show that after in vivo delivery the rAAV-rDNA vectors integrated into the genomic rDNA locus 8-13 times more frequently than control vectors, providing an estimate that 23-39% of the integrations were specific to the rDNA locus. Moreover, a rAAV-rDNA vector containing a human factor IX (hFIX) expression cassette resulted in sustained therapeutic levels of serum hFIX even after repeated manipulations to induce liver regeneration. Because of the relative safety of integration in the rDNA locus, these vectors expand the usage of rAAV for therapeutics requiring long-term gene transfer into dividing cells.

Original languageEnglish (US)
Pages (from-to)1912-1923
Number of pages12
JournalMolecular Therapy
Volume20
Issue number10
DOIs
StatePublished - Oct 2012

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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    Lisowski, L., Lau, A., Wang, Z., Zhang, Y., Zhang, F., Grompe, M., & Kay, M. A. (2012). Ribosomal DNA integrating rAAV-rDNA vectors allow for stable transgene expression. Molecular Therapy, 20(10), 1912-1923. https://doi.org/10.1038/mt.2012.164