Abstract
Virus-induced gene silencing (VIGS) uses recombinant viruses to study gene function; however, the effect of the virus vector itself on the gene expression of the host is not always considered. In our work, we investigated non-targeted gene expression changes of the host in order to see how often these changes appear. Effects of various VIGS vector infections were analysed by monitoring gene expression levels of housekeeping genes by Northern blot analysis in four different hosts. We found that non-targeted changes happens very often. More importantly, these non-targeted effects can cause drastic changes in the gene-expression pattern of host genes that are usually used as references in these studies. We have also found that in a tobacco rattle virus (TRV)-based VIGS, the presence of foreign sequences in the cloning site of the vector can also have a non-targeted effect, and even the use of an internal control can lead to unpredicted changes. Our results show that although VIGS is a very powerful technique, the VIGS vector, as a pathogen of the host, can cause unwanted changes in its gene-expression pattern, highlighting the importance of careful selection of both the genes to be tested and those to be used as references in the planned experiments.
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Acknowledgements
The authors are grateful to C. Lacomme (University of Edinburgh) for BSMV and TRV, and to D. Baulcombe (University of Cambridge, UK) for TRV, PVX and TMV vectors. The TRV2-Upf1 vector was kindly provided by Dániel Silhavy. This work was funded by OTKA Grants K108718 and K109438. EO is a student of the Plant Science Doctoral School at Szent István University, Gödöllő. RP is a student of the Doctoral School of Biological Sciences at Szent István University, Gödöllő. DT is a student of the Doctoral School of Crop and Horticultural Sciences at Pannon University, Keszthely. We thank Matyas Medzichardszky for careful reading of our manuscript.
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Oláh, E., Pesti, R., Taller, D. et al. Non-targeted effects of virus-induced gene silencing vectors on host endogenous gene expression. Arch Virol 161, 2387–2393 (2016). https://doi.org/10.1007/s00705-016-2921-9
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DOI: https://doi.org/10.1007/s00705-016-2921-9