Plant RNA Silencing in Viral Defence

Pantaleo, Vitantonio

doi:10.1007/978-1-4614-0332-6_3

Cited by 34 publications

(17 citation statements)

References 135 publications

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“…We tested two virus platforms because cr-TMV/TVCV can spread only by cell-to-cell movement in N. bentamiana , whereas PVX can also spread systemically [35], [36], [52]. The resulting fluorescence assays (Fig.…”

Section: Resultsmentioning

confidence: 99%

High-Level Transient Expression of ER-Targeted Human Interleukin 6 in Nicotiana benthamiana

et al. 2012

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Tobacco plants can be used to express recombinant proteins that cannot be produced in a soluble and active form using traditional platforms such as Escherichia coli. We therefore expressed the human glycoprotein interleukin 6 (IL6) in two commercial tobacco cultivars (Nicotiana tabacum cv. Virginia and cv. Geudertheimer) as well as the model host N. benthamiana to compare different transformation strategies (stable vs. transient expression) and subcellular targeting (apoplast, endoplasmic reticulum (ER) and vacuole). In T0 transgenic plants, the highest expression levels were achieved by ER targeting but the overall yields of IL6 were still low in the leaves (0.005% TSP in the ER, 0.0008% in the vacuole and 0.0005% in the apoplast). The apoplast variant accumulated to similar levels in leaves and seeds, whereas the ER-targeted variant was 1.2-fold more abundant in seeds and the vacuolar variant was 6-fold more abundant in seeds. The yields improved in subsequent generations, with the best-performing T2 plants producing the ER-targeted IL6 at 0.14% TSP in both leaves and seeds. Transient expression of ER-targeted IL6 in leaves using the MagnICON system resulted in yields of up to 7% TSP in N. benthamiana, but only 1% in N. tabacum cv. Virginia and 0.5% in cv. Geudertheimer. Although the commercial tobacco cultivars produced up to threefold more biomass than N. benthamiana, this was not enough to compensate for the lower overall yields. The recombinant IL6 produced by transient and stable expression in plants was biologically active and presented as two alternative bands matching the corresponding native protein.

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Section: Resultsmentioning

confidence: 99%

High-Level Transient Expression of ER-Targeted Human Interleukin 6 in Nicotiana benthamiana

et al. 2012

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“…The RNAi response is a sequence specific breakdown mechanism, which can be divided into different pathways dependent on the small RNA molecules involved. The small interfering (si)RNA pathway is known to act antivirally in plants and arthropods [102,103]. It is induced by exogenous long dsRNA molecules that are recognized and cleaved into siRNA molecules.…”

Section: Genus Tospovirusmentioning

confidence: 99%

Non-Structural Proteins of Arthropod-Borne Bunyaviruses: Roles and Functions

Eifan

Schnettler

Dietrich

et al. 2013

Viruses

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Viruses within the Bunyaviridae family are tri-segmented, negative-stranded RNA viruses. The family includes several emerging and re-emerging viruses of humans, animals and plants, such as Rift Valley fever virus, Crimean-Congo hemorrhagic fever virus, La Crosse virus, Schmallenberg virus and tomato spotted wilt virus. Many bunyaviruses are arthropod-borne, so-called arboviruses. Depending on the genus, bunyaviruses encode, in addition to the RNA-dependent RNA polymerase and the different structural proteins, one or several non-structural proteins. These non-structural proteins are not always essential for virus growth and replication but can play an important role in viral pathogenesis through their interaction with the host innate immune system. In this review, we will summarize current knowledge and understanding of insect-borne bunyavirus non-structural protein function(s) in vertebrate, plant and arthropod.

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“…What makes this mechanism of great interest is that the gene silencing is highly specific, and also highly potent in where only a few copies of an small RNA (22–25 nt long) can demonstrate wide ranging affects. In plants, they have been highly investigated for their role in virus response (Ridanpaa et al, 2003; Pantaleo, 2011), but in humans and mice they are under intense study for therapeutic medicine (recently reviewed in Burnette et al, 2005; Khaliq et al, 2010; Vaishnaw et al, 2011). In an example, a combination of host and viral genes has been used as a siRNA-based treatment for hepatitis C virus (HCV; Ashfaq et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Characterizing ncRNAs in human pathogenic protists using high-throughput sequencing technology

Collins¹

2011

fgene

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ncRNAs are key genes in many human diseases including cancer and viral infection, as well as providing critical functions in pathogenic organisms such as fungi, bacteria, viruses, and protists. Until now the identification and characterization of ncRNAs associated with disease has been slow or inaccurate requiring many years of testing to understand complicated RNA and protein gene relationships. High-throughput sequencing now offers the opportunity to characterize miRNAs, siRNAs, small nucleolar RNAs (snoRNAs), and long ncRNAs on a genomic scale, making it faster and easier to clarify how these ncRNAs contribute to the disease state. However, this technology is still relatively new, and ncRNA discovery is not an application of high priority for streamlined bioinformatics. Here we summarize background concepts and practical approaches for ncRNA analysis using high-throughput sequencing, and how it relates to understanding human disease. As a case study, we focus on the parasitic protists Giardia lamblia and Trichomonas vaginalis, where large evolutionary distance has meant difficulties in comparing ncRNAs with those from model eukaryotes. A combination of biological, computational, and sequencing approaches has enabled easier classification of ncRNA classes such as snoRNAs, but has also aided the identification of novel classes. It is hoped that a higher level of understanding of ncRNA expression and interaction may aid in the development of less harsh treatment for protist-based diseases.

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Plant RNA Silencing in Viral Defence

Cited by 34 publications

References 135 publications

High-Level Transient Expression of ER-Targeted Human Interleukin 6 in Nicotiana benthamiana

High-Level Transient Expression of ER-Targeted Human Interleukin 6 in Nicotiana benthamiana

Non-Structural Proteins of Arthropod-Borne Bunyaviruses: Roles and Functions

Characterizing ncRNAs in human pathogenic protists using high-throughput sequencing technology

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