Non-targeted effects of virus-induced gene silencing vectors on host endogenous gene expression

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.

Crispr/Cas9 Mediated Inactivation of Argonaute 2 Reveals its Differential Involvement in Antiviral Responses

“…Viral vectors applied for VIGS experiments (TRV, PVX etc.) may also interfere with various endogenous cellular functions and cause unintended changes in the expression of non-target genes, making it difficult to unambiguously interpret the results 59 . This can become especially problematic for studying host-virus interactions, since viruses often exhibit synergistic or antagonistic relationships with each other.…”

Virus-Induced Gene Silencing (VIGS): A Powerful Tool for Crop Improvement and Its Advancement towards Epigenetics

“…First, the virus vector can disrupt plant metabolism [ 116 , 117 ], affecting plant–microbe interaction. The insertion of a gene into the VIGS vector can prevent the virus from multiplying, and several viruses have been known to delete the inserted gene during multiplication and spread [ 118 ].…”

Analysis of a novel RNA virus in a wild northern white-breasted hedgehog (Erinaceus roumanicus)

“…During the plant inoculation study, Nicotiana benthamiana , Chenopodium quinoa and C. murale plants were grown at 21-22 °C. Eight- to ten-week-old plants with four leaves were leaf-rub inoculated with transcripts containing or not containing (mock) inoculation buffer [16]. Infected plants were kept at 21-22 °C in a growth chamber with a 16-h light/8-h dark cycle.…”