Nucleic Acid Sensing in Invertebrate Antiviral Immunity

Wang, Pei‐Hui; He, Jianguo

doi:10.1016/bs.ircmb.2018.11.002

Cited by 32 publications

(26 citation statements)

References 208 publications

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“…SID-1 proteins first identified in C. elegans allow passive transport of dsRNA into the cell and are found ubiquitously in metazoan evolution, however whether they contribute to systemic RNAi seems to vary [18,19]. In crustaceans, however, SID-1 proteins in Macrobrachium rosenbergii and Litopenaeus vannamei were shown to facilitate dsRNA uptake [20][21][22]. Uptake of dsRNA may also occur via receptor-mediated endocytosis, as has been demonstrated in C. elegans and some insect species [17,[23][24][25].…”

Section: Known Mechanisms Of Systemic Rnai-a Drop In the Oceanmentioning

confidence: 98%

Double-Stranded RNA Binding Proteins in Serum Contribute to Systemic RNAi Across Phyla—Towards Finding the Missing Link in Achelata

Banks

Wang

Fitzgibbon

et al. 2020

IJMS

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RNA interference (RNAi) has become a widely utilized method for studying gene function, yet despite this many of the mechanisms surrounding RNAi remain elusive. The core RNAi machinery is relatively well understood, however many of the systemic mechanisms, particularly double-stranded RNA (dsRNA) transport, are not. Here, we demonstrate that dsRNA binding proteins in the serum contribute to systemic RNAi and may be the limiting factor in RNAi capacity for species such as spiny lobsters, where gene silencing is not functional. Incubating sera from a variety of species across phyla with dsRNA led to a gel mobility shift in species in which systemic RNAi has been observed, with this response being absent in species in which systemic RNAi has never been observed. Proteomic analysis suggested lipoproteins may be responsible for this phenomenon and may transport dsRNA to spread the RNAi signal systemically. Following this, we identified the same gel shift in the slipper lobster Thenus australiensis and subsequently silenced the insulin androgenic gland hormone, marking the first time RNAi has been performed in any lobster species. These results pave the way for inducing RNAi in spiny lobsters and for a better understanding of the mechanisms of systemic RNAi in Crustacea, as well as across phyla.

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Section: Known Mechanisms Of Systemic Rnai-a Drop In the Oceanmentioning

confidence: 98%

Double-Stranded RNA Binding Proteins in Serum Contribute to Systemic RNAi Across Phyla—Towards Finding the Missing Link in Achelata

Banks

Wang

Fitzgibbon

et al. 2020

IJMS

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“…LvIRF can be activated during viral infection, and then translocates to the nucleus to initiate the expression of the L. vannamei Vago4 (LvVago4) gene. This is effectively an arthropod cytokine encoding a viral-activated secreted peptide through activating the JAK-STAT pathway to restrict viral infection, similar to mammalian IFNs (Chen et al, 2011;Paradkar et al, 2012;Wang and He, 2019).…”

Section: Introductionmentioning

confidence: 99%

A Polymorphic (CT)n-SSR Influences the Activity of the Litopenaeus vannamei IRF Gene Implicated in Viral Resistance

et al. 2019

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Simple sequence repeats (SSRs) of short nucleotide motifs occur very frequently in the 5′ untranslated coding region (5′-UTR) of genes and have been implicated in the regulation of gene expression. In this study, we identified an SSR with a variable number of CT repeats in the 5′-UTR of the Litopenaeus vannamei IRF (LvIRF) gene that has been shown to mediate antiviral responses by inducing the expression of Vago, a functional homolog of mammalian IFN. We then explored the effects of varying the number of (CT)n repeats on the expression of LvIRF using both dual-luciferase reporter assays and Western blots. Our results demonstrate that the length of the (CT)n-SSR in this gene can influence the expressional level of LvIRF, in that a shorter (CT)n repeat had a stronger ability to induce the expression of LvIRF. Moreover, we found that the (CT)n repeat in LvIRF was associated with viral resistance in shrimp. Individual shrimps with shorter (CT)n repeats in the 5′-UTR of LvIRF exhibited high tolerance to white spot syndrome virus (WSSV), and this trait was inherited in offspring. Taken together, these results indicated that this (CT)n-SSR could be used as a molecular marker for shrimp breeding for WSSV resistance.

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“…SID-1 proteins first identified in C. elegans, allow passive transport of dsRNA into the cell and are found ubiquitously in metazoan evolution, however whether they contribute to systemic RNAi seems to vary [19,20]. In crustaceans however, SID-1 proteins in Macrobrachium rosenbergii and Litopenaeus vannamei were shown to facilitate dsRNA uptake [21][22][23]. Uptake of dsRNA may also occur via receptor mediated endocytosis, as has been demonstrated in C. elegans and some insect species [18,[24][25][26].…”

Section: Known Mechanisms Of Systemic Rnai-a Drop In the Oceanmentioning

confidence: 99%

Double Stranded RNA Binding Proteins in the Serum Contribute to Systemic RNAi across Phyla – Towards Finding the Missing Link in Achelata

Banks¹,

Wang²,

Fitzgibbon³

et al. 2020

Preprint

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RNA interference (RNAi) has become a widely utilised method to study gene function, yet despite this, many of the mechanisms surrounding RNAi remain elusive. The core RNAi machinery is relatively well understood, however many of the systemic mechanisms, particularly double stranded RNA (dsRNA) transport, are not. Here, we demonstrate that dsRNA binding proteins in the serum contribute to systemic RNAi, and may be the limiting factor in RNAi capacity for species such as spiny lobsters where gene silencing is not functional. Incubating serum from a variety of species across phyla with dsRNA led to a gel mobility shift in species where systemic RNAi has been observed, with this response being absent in species where systemic RNAi has never been observed. Proteomic analysis suggested lipoproteins may be responsible for this phenomenon, and may transport dsRNA to spread the RNAi signal systemically. Following this, we identified the same gel shift in the slipper lobster Thenus australiensis and subsequently silenced the insulin androgenic gland hormone, marking the first time RNAi has been performed in any lobster species. These results pave the way for inducing RNAi in spiny lobsters, and better understanding the mechanisms of systemic RNAi in Crustacea, as well as across phyla.

show abstract

Nucleic Acid Sensing in Invertebrate Antiviral Immunity

Cited by 32 publications

References 208 publications

Double-Stranded RNA Binding Proteins in Serum Contribute to Systemic RNAi Across Phyla—Towards Finding the Missing Link in Achelata

Double-Stranded RNA Binding Proteins in Serum Contribute to Systemic RNAi Across Phyla—Towards Finding the Missing Link in Achelata

A Polymorphic (CT)n-SSR Influences the Activity of the Litopenaeus vannamei IRF Gene Implicated in Viral Resistance

Double Stranded RNA Binding Proteins in the Serum Contribute to Systemic RNAi across Phyla – Towards Finding the Missing Link in Achelata

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