Transposition from a gutless adeno-transposon vector stabilizes transgene expression in vivo

Abstract

A major limitation of adenovirus-mediated gene therapy for inherited diseases is the instability of transgene expression in vivo, which originates at least in part from the loss of the linear, extrachromosomal vector genomes. Herein we describe the production of a gene-deleted adenovirus-transposon vector that stably maintains virus-encoded transgenes in vivo through integration into host cell chromosomes. This system utilizes a donor transposon vector that undergoes Flp-mediated recombination and excision of its therapeutic payload in the presence of the Flp and Sleeping Beauty recombinases. Systemic in vivo delivery of this system resulted in efficient generation of transposon circles and stable transposase-mediated integration in mouse liver. Somatic integration was sufficient to maintain therapeutic levels of human coagulation Factor IX for more than six months in mice undergoing extensive liver proliferation. These vectors combine the versatility of adenoviral vectors with the integration capabilities of a eukaryotic DNA transposon and should prove useful in the treatment of genetic diseases.

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

“…24 Conversely, the SB system comprises the SB transposase and the genetic cargo framed by the terminal inverted repeats/directrepeat (IRs/DR), whereby the SB transposase binding to the IR/DR sequences results in the transposon excision from the donor and subsequent integration into the chromosome (cut-and-paste mechanism). 25 The SB system has been used in conjunction with adenovirus 26 or herpes simplex virus amplicon 27 to generate hybrid vectors for prolonged expression.…”

Viral vectors for in vivo gene transfer in Parkinson's disease: Properties and clinical grade production

“…In general, rAd and HSV do not integrate into the genome and form episomes (Harui et al, 1999;Hillgenberg et al, 2001;Oehmig et al, 2004b). Both rAd and HSV can be engineered to integrate into the genome, thus, addition of a transposon to a helper-dependent (high capacity) rAd, increases the frequency of integration and enhances long-term gene expression (Yant et al, 2002). Likewise, HSV amplicon vectors can be induced to integrate by including sequences from viruses that normally integrate into the genome (Oehmig et al, 2004b).…”

Gutless adenovirus: last-generation adenovirus for gene therapy

“…110,111 Other tissues like lung, cardiac muscle, vascular tissue or dendritic cells have also been targeted with gutless adenoviruses with similar results as for the muscle or the liver. [112][113][114][115] Future considerations This is a fascinating moment for gutless adenovirus vectors: their use permits long-term expression in vivo with reduced and transient cellular immune response in Gutless adenovirus R Alba et al animal models for human diseases and, moreover, the increasing number of groups working in this field favors technological advances to escape preimmune response by developing non-human gutless adenovirus 20 or to increase stability by generating integrative gutless vectors, 116,117 or vectors with replication capacity. 118 However, their use in clinical assays is questionable since helper contamination levels are still high, and large-scale production in bioreactors is not yet fully developed.…”