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| Item Type: | Article |
|---|---|
| Title: | Integration profile and safety of an adenovirus hybrid-vector utilizing hyperactive Sleeping Beauty transposase for somatic integration |
| Creators Name: | Zhang, W., Muck-Hausl, M., Wang, J., Sun, C., Gebbing, M., Miskey, C., Ivics, Z., Izsvak, Z. and Ehrhardt, A. |
| Abstract: | We recently developed adenovirus/transposase hybrid-vectors utilizing the previously described hyperactive Sleeping Beauty (SB) transposase HSB5 for somatic integration and we could show stabilized transgene expression in mice and a canine model for hemophilia B. However, the safety profile of these hybrid-vectors with respect to vector dose and genotoxicity remains to be investigated. Herein, we evaluated this hybrid-vector system in C57Bl/6 mice with escalating vector dose settings. We found that in all mice which received the hyperactive SB transposase, transgene expression levels were stabilized in a dose-dependent manner and that the highest vector dose was accompanied by fatalities in mice. To analyze potential genotoxic side-effects due to somatic integration into host chromosomes, we performed a genome-wide integration site analysis using linker-mediated PCR (LM-PCR) and linear amplification-mediated PCR (LAM-PCR). Analysis of genomic DNA samples obtained from HSB5 treated female and male mice revealed a total of 1327 unique transposition events. Overall the chromosomal distribution pattern was close-to-random and we observed a random integration profile with respect to integration into gene and non-gene areas. Notably, when using the LM-PCR protocol, 27 extra-chromosomal integration events were identified, most likely caused by transposon excision and subsequent transposition into the delivered adenoviral vector genome. In total, this study provides a careful evaluation of the safety profile of adenovirus/Sleeping Beauty transposase hybrid-vectors. The obtained information will be useful when designing future preclinical studies utilizing hybrid-vectors in small and large animal models. |
| Keywords: | Adenoviridae, Computational Biology, Enzyme-Linked Immunosorbent Assay, Genetic Vectors, HeLa Cells, Transposases, Animals, Mice |
| Source: | PLoS ONE |
| ISSN: | 1932-6203 |
| Publisher: | Public Library of Science |
| Volume: | 8 |
| Number: | 10 |
| Page Range: | e75344 |
| Date: | 4 October 2013 |
| Additional Information: | Erratum in: PLoS One. 2020 Feb 4;15(2):e0228902. |
| Official Publication: | https://doi.org/10.1371/journal.pone.0075344 |
| PubMed: | View item in PubMed |
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