Helmholtz Gemeinschaft

Search
Browse
Statistics
Feeds

Integration profile and safety of an adenovirus hybrid-vector utilizing hyperactive Sleeping Beauty transposase for somatic integration

[img] PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
1MB

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. and Muck-Hausl, M. and Wang, J. and Sun, C. and Gebbing, M. and Miskey, C. and Ivics, Z. and 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 (U.S.A.)
Volume:8
Number:10
Page Range:e75344
Date:4 October 2013
Official Publication:https://doi.org/10.1371/journal.pone.0075344
PubMed:View item in PubMed

Repository Staff Only: item control page

Downloads

Downloads per month over past year

Open Access
MDC Library