We first established that increasing the number of IRESmediated
gene cassettes increases corresponding gene expression in our Nicotiana benthamiana transient expression assay (Figure 2). As an application of this system, we also demonstrated that expression of the Tombusvirus p19 protein, an RNA silencing suppressor, from a double IRES cassette—but not single cassette— suppressed RNA silencing of its own mRNA (Figure 3). This is the first report of a mRNA self-protection system from RNA silencing by IRES-mediated expression of a Figure 4. Western blot analysis of Venus and HA-tagged PYR1 transient expression from vector, PYR1, PIV and PI⊿V. RuBisCo protein was used as a loading control. Copyright © 2015 The Japanese Society for Plant Cell and Molecular Biology viral suppressor gene in plants. Here, we propose that it is beneficial to use polycistronic expression of RNA silencing suppressors as protection for a plant’s own mRNA.
We first established that increasing the number of IRESmediatedgene cassettes increases corresponding gene expression in our Nicotiana benthamiana transient expression assay (Figure 2). As an application of this system, we also demonstrated that expression of the Tombusvirus p19 protein, an RNA silencing suppressor, from a double IRES cassette—but not single cassette— suppressed RNA silencing of its own mRNA (Figure 3). This is the first report of a mRNA self-protection system from RNA silencing by IRES-mediated expression of a Figure 4. Western blot analysis of Venus and HA-tagged PYR1 transient expression from vector, PYR1, PIV and PI⊿V. RuBisCo protein was used as a loading control. Copyright © 2015 The Japanese Society for Plant Cell and Molecular Biology viral suppressor gene in plants. Here, we propose that it is beneficial to use polycistronic expression of RNA silencing suppressors as protection for a plant’s own mRNA.
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