• Korean journal of applied entomology
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• v.55 no.2
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• pp.81-89
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• 2016

Double-stranded RNA (dsRNA) has been applied to control insect pests by its suppressive activity against specific target genes. Integrin is a heterodimer (${\alpha}$ and ${\beta}$) transmembrane protein and plays a critical role in cell-to-cell or cell-to-extracellular matrix interactions in eukaryotes. Suppression of ${\beta}$ subunit integrin gene expression by its specific dsRNA (= dsINT) induces significant mortality against target insects. Furthermore, a recombinant bacterium expressing dsINT is potent to kill target insects. However, it is necessary to develop a formulation technique of the dsRNA-expressing bacteria to apply the bacterial insecticide against field populations. This study formulated the recombinant bacteria by freeze-drying and tested its control efficacy against target insects. The formulation maintained significant insecticidal activity against last instar larvae of Spodoptera exigua. While a commercial Bacillus thuringiensis (Bt) insecticide exhibited only about 60% insecticidal activity against S. exigua last instar, an addition of the dsINT-expressing bacterial formulation significantly enhanced the Bt insecticidal activity. The dsINT-expressing bacterial formulation exhibited relative selectivity to target insects depending on sequence similarity. These results indicate that a freeze-dried form of dsRNA-expressing bacteria keeps its insecticidal activity.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.460-464
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• 2013

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed identical sequences sequence to known RNA-dependent RNA polymerase genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.202-209
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• 2014

RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

• Korean Journal of Microbiology
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• v.33 no.2
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• pp.105-110
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• 1997

Ustilago maydis strains (A-series and SH-series) containg virus or viral dsRNAs were artificially mated in corn seedling to generate 6 progeny strains, designated A23, A45, A21l, A31O, SH24 and SH61O. The dsRNA patterns of progeny strains were identical to those of the parental strains and there was no molecular exclusion mechanism among dsRNAs of parental strains. Virus particles were purified from 6 progeny strains and viral dsRNAs were analyzed on 5% PAGE. There was no mixed encapsidation between virus or dsRNAs of parental strains. Progeny strain SH6l4 produced toxin which inhibits the growth of SH9, SHIO and SH11. Likewise, toxins from A310 and SH24 inhibited growth of the SH11 strains. These results indicate that the presence of different types of dsRNA does not interfere the expression of toxin gene.

• Fisheries and Aquatic Sciences
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• v.13 no.1
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• pp.56-62
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• 2010

To determine whether long double-stranded RNA (dsRNA) induces RNA interference and type I interferon (IFN) responses in fish, long dsRNAs encoding enhanced green fluorescent protein (EGFP), GFPuv, and polyinosinic-polycytidylic acid sequences were co-injected with an EGFP expressing plasmid, into rock bream (Oplegnathus fasciatus). We investigated the EGFP mRNA and protein levels, and the transcriptional responses of dsRNA-dependent protein kinase and Mx1 genes. Long dsRNAs were strong inducers of a type I IFN response in rock bream, resulting in nonspecific suppression of exogenous gene expression. Furthermore, sequence-specific knockdown of exogenous gene expression at the mRNA level was detected at an early phase (24 h). These results suggested that long dsRNA may inhibit exogenous gene expression through an early mRNA interference response and a later type I IFN response in fish.

• Research in Plant Disease
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• v.12 no.1
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• pp.62-64
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• 2006

July in 2005, we collected infected maize plant that showing stripe dwarf disease on maize leaf in Jeonbuk provinces including Gochang-gun and conducted genomic dsRNA extraction and RT-PCR. Genomic dsRNA was extracted directly in infected maize plant and electrophoresis in agarose gel. We confirmed 10 segments of genomic dsRNA. We conducted RT-PCR by genomic dsRNA and specific primer of S7, S8 and S10. As a result, specific band of expected size was confirmed respectively. In the results of dsRNA and RT-PCR analysis, we confirmed Rice black-streaked dwarf fijivirus (RBSDV) from naturally infected maize plant. Occurrence of RBSDV of maize plant was dealt 22 ha's damage in maize field. The occurrence rate was 80% in a lot of places of disease.

• Korean Journal of Microbiology
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• v.31 no.3
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• pp.184-188
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• 1993

Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

• Applied Biological Chemistry
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• v.34 no.2
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• pp.86-93
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• 1991

cis-Diamminedichloroplatinum(II)(CDDP), an antitumor drug, did not generate crosslink between bluetongue virus (BTV) capsid protein at moderate concentration. Cesium chloride density gradient centrifugation study revealed that protein-RNA crosslink was not detectable in CDDP treated BTV. CDDP treated BTV ds RNA showed remarkable change in the migration pattern in polyacrylamide gel electrophoresis. These results suggest that the reduction of BTV core associated transcriptase activity is most likely by the CDDP adduction to the genomic ds RNA rather than by the protein-RNA crosslink and/or protein-protein cross-link.

• IMMUNE NETWORK
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• v.22 no.2
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• pp.15.1-15.16
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• 2022

Foreign molecules, including viruses and bacteria-derived toxins, can also induce airway inflammation. However, to the best of our knowledge, the roles of these molecules in the development of airway inflammation have not been fully elucidated. Herein, we investigated the precise role and synergistic effect of virus-mimicking double-stranded RNA (dsRNA) and staphylococcal enterotoxin B (SEB) in macrophages and epithelial cells. To identify cytokine expression profiles, both the THP-1-derived macrophages and BEAS-2B epithelial cells were stimulated with dsRNA or SEB. A total of 21 cytokines were evaluated in the culture supernatants. We observed that stimulation with dsRNA induced cytokine production in both cell types. However, cytokine production was not induced in SEB-stimulated epithelial cells, compared to the macrophages. The synergistic effect of dsRNA and SEB was evaluated observing cytokine level and intracellular phospho-signaling. Fifteen different types were detected in high-dose dsRNA-stimulated epithelial cells, and 12 distinct types were detected in macrophages; those found in macrophages lacked interferon production compared to the epithelial cells. Notably, a synergistic effect of cytokine induction by co-stimulation of dsRNA and SEB was observed mainly in epithelial cells, via activation of most intracellular phosphor-signaling. However, macrophages only showed an accumulative effect. This study showed that the type and severity of cytokine productions from the epithelium or macrophages could be affected by different intensities and a combination of dsRNA and SEB. Further studies with this approach may improve our understanding of the development and exacerbation of airway inflammation and asthma.

• Korean journal of applied entomology
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• v.61 no.1
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• pp.211-219
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• 2022

Over the past decade, double-stranded RNA (dsRNA)-mediated gene silencing technology has progressed significantly for pest management in agriculture and for protecting beneficial insects from pathogens. Recently, breakthroughs in RNA interference (RNAi) applications for insect pest management by academia and commercial entities have provided RNAi products as commercial biopesticides. Although RNAi technology has vast potential and advantages for pest control, challenges, and limitations remain in practical applications. This review explores current challenges in the development of dsRNAs as a pest management tool and considers new approaches to overcome biological and environmental obstacles, such as poor stability and resistance.