E-RNAi: a web application for the multi-species design of RNAi reagents—2010 update

Horn, Thomas; Boutros, Michael

doi:10.1093/nar/gkq317

Cited by 144 publications

(105 citation statements)

References 25 publications

(35 reference statements)

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“…Primers were designed from the sequence of the A. mellifera nkd gene (GenBank accession no. XM_001120899) by using the E-RNAi web service (56), and primers for GFP, which served as the control gene, were used from previous studies (57). All primer sequences were fused with the T7 promoter sequence (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Silencing the Honey Bee (Apis mellifera) Naked Cuticle Gene ( nkd ) Improves Host Immune Function and Reduces Nosema ceranae Infections

Li¹,

Evans²,

Huang

et al. 2016

Appl Environ Microbiol

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Nosema ceranae is a new and emerging microsporidian parasite of European honey bees, Apis mellifera, that has been implicated in colony losses worldwide. RNA interference (RNAi), a posttranscriptional gene silencing mechanism, has emerged as a potent and specific strategy for controlling infections of parasites and pathogens in honey bees. While previous studies have focused on the silencing of parasite/pathogen virulence factors, we explore here the possibility of silencing a host factor as a mechanism for reducing parasite load. Specifically, we used an RNAi strategy to reduce the expression of a honey bee gene, naked cuticle (nkd), which is a negative regulator of host immune function. Our studies found that nkd mRNA levels in adult bees were upregulated by N. ceranae infection (and thus, the parasite may use this mechanism to suppress host immune function) and that ingestion of double-stranded RNA (dsRNA) specific to nkd efficiently silenced its expression. Furthermore, we found that RNAimediated knockdown of nkd transcripts in Nosema-infected bees resulted in upregulation of the expression of several immune genes (Abaecin, Apidaecin, Defensin-1, and PGRP-S2), reduction of Nosema spore loads, and extension of honey bee life span. The results of our studies clearly indicate that silencing the host nkd gene can activate honey bee immune responses, suppress the reproduction of N. ceranae, and improve the overall health of honey bees. This study represents a novel host-derived therapeutic for honey bee disease treatment that merits further exploration. IMPORTANCEGiven the critical role of honey bees in the pollination of agricultural crops, it is urgent to develop strategies to prevent the colony decline induced by the infection of parasites/pathogens. Targeting parasites and pathogens directly by RNAi has been proven to be useful for controlling infections in honey bees, but little is known about the disease impacts of RNAi silencing of host factors. Here, we demonstrate that knocking down the honey bee immune repressor-encoding nkd gene can suppress the reproduction of N. ceranae and improve the overall health of honey bees, which highlights the potential role of host-derived and RNAi-based therapeutics in controlling the infections in honey bees. The information obtained from this study will have positive implications for honey bee disease management practices. E uropean honey bees, Apis mellifera, play a critical role in the pollination of important crops. However, honey bee populations have suffered high losses in much of the world (1), coincident with an increase in agricultural demand for honey bee pollination (2). Specifically, honey bee colony losses in the United States have been exacerbated since the report of colony collapse disorder (CCD), a syndrome that comprises large-scale, unexplained losses of managed honey bees (3-9). High levels of parasites and pathogens have been linked to the decline of honey bee colonies (10,11).Nosema is a genus of obligate, intracellular microsporidian parasite...

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Section: Methodsmentioning

confidence: 99%

Silencing the Honey Bee (Apis mellifera) Naked Cuticle Gene ( nkd ) Improves Host Immune Function and Reduces Nosema ceranae Infections

Li¹,

Evans²,

Huang

et al. 2016

Appl Environ Microbiol

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“…dsRNAs were prepared by in vitro transcription (MEGAscript RNAi, Ambion) of either PCR products or cDNAs cloned in the pBluescript vector. Amplified sequences were selected with the E-RNAi software (26).…”

Section: Methodsmentioning

confidence: 99%

dTIS11 Protein-dependent Polysomal Deadenylation Is the Key Step in AU-rich Element-mediated mRNA Decay in Drosophila Cells

Vindry

Lauwers

Hutin

et al. 2012

Journal of Biological Chemistry

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Background: TIS11 proteins control the degradation of mRNA containing AU-rich elements (ARE). Results: Drosophila dTIS11 activates the polysomal deadenylation of the ARE mRNA Cecropin A1. Conclusion: Drosophila dTIS11 controls fewer aspects of ARE mRNA decay as compared with its mammalian homologs. Significance: TIS11 proteins may have acquired additional functions to control mRNA decay during evolution from invertebrates to mammals.

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“…Small interfering RNAs (siRNAs) directed against the DIS gene transcript were designed using version 3.2 of E-RNAi (Horn and Boutros, 2010). The specificity of the designed siRNAs was determined by comparing the sequence (BLASTn) with complete genome and transcriptome sequences.…”

Section: Rnai Knockdown Of Dis Expressionmentioning

confidence: 99%

DISTAG/TBCCd1 Is Required for Basal Cell Fate Determination in Ectocarpus

et al. 2017

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Brown algae are one of the most developmentally complex groups within the eukaryotes. As in many land plants and animals, their main body axis is established early in development, when the initial cell gives rise to two daughter cells that have apical and basal identities, equivalent to shoot and root identities in land plants, respectively. We show here that mutations in the Ectocarpus DISTAG (DIS) gene lead to loss of basal structures during both the gametophyte and the sporophyte generations. Several abnormalities were observed in the germinating initial cell in dis mutants, including increased cell size, disorganization of the Golgi apparatus, disruption of the microtubule network, and aberrant positioning of the nucleus. DIS encodes a TBCCd1 protein, which has a role in internal cell organization in animals, Chlamydomonas reinhardtii, and trypanosomes. Our study highlights the key role of subcellular events within the germinating initial cell in the determination of apical/basal cell identities in a brown alga and emphasizes the remarkable functional conservation of TBCCd1 in regulating internal cell organization across extremely distant eukaryotic groups.

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E-RNAi: a web application for the multi-species design of RNAi reagents—2010 update

Cited by 144 publications

References 25 publications

Silencing the Honey Bee (Apis mellifera) Naked Cuticle Gene ( nkd ) Improves Host Immune Function and Reduces Nosema ceranae Infections

Silencing the Honey Bee (Apis mellifera) Naked Cuticle Gene ( nkd ) Improves Host Immune Function and Reduces Nosema ceranae Infections

dTIS11 Protein-dependent Polysomal Deadenylation Is the Key Step in AU-rich Element-mediated mRNA Decay in Drosophila Cells

DISTAG/TBCCd1 Is Required for Basal Cell Fate Determination in Ectocarpus

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