Inhibition of BmNPV replication in silkworm cells using inducible and regulated artificial microRNA precursors targeting the essential viral gene lef-11

Zhang, Jun; He, Qian; Zhang, Chundong; Chen, Xiangyun; Chen, Xuemei; Dong, Zhanqi; Li, Na; Kuang, Xiu-Xiu; Cao, Mengji; Lü, Cheng; Pan, Min‐Hui

doi:10.1016/j.antiviral.2014.01.017

Cited by 44 publications

(37 citation statements)

References 35 publications

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“…However, silkworm disease caused annually by BmNPV results in huge economic losses in the sericulture industry (Jiang and Xia, ). RNAi and overexpression of extraneous proteins have been used to generate transgenic antiviral silkworm lines as a potential therapeutic approach (Zhang, He et al , ; Dong et al , ). However, it is difficult to significantly increase the antiviral effect, which hinders its application in sericulture and in conducting further in‐depth studies.…”

Section: Discussionmentioning

confidence: 99%

“…The majority of previous antiviral studies have reported the use of knockdown of baculovirus DNA replication factor immediate early‐1 (IE‐1) gene through RNAi technology, which significantly inhibited the virus replication in vivo and in vitro (Zhang, Wang et al , ; Zhou et al , ). Moreover, ie‐1 , lef‐1 , lef‐2 , lef‐3 , lef‐11 , oral infection factor p74 and gp64 have also significantly inhibited virus multiplication and replication in transgenic cells and silkworm (Fujita et al , ; Jiang et al , ; Xu et al , ; Zhang, He et al , ; Zhou et al , ; Cao et al , ). However, baculovirus early transcription activator and DNA replication factors, ie‐0 and ie‐2 , have not yet been studied as target genes.…”

Section: Introductionmentioning

confidence: 99%

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CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

Dong

et al. 2018

Insect Molecular Biology

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The CRISPR/Cas9 system is a powerful tool for the treatment of infectious diseases. In our previous study, we knocked out the Bombyx mori nucleopolyhedrovirus (BmNPV) key genes and BmNPV-dependent host factor to generate transgenic antiviral strains. To further expand the range of target genes for BmNPV and more effectively prevent and control pathogenic infections, we performed gene editing and antiviral analysis by constructing a target-directed baculovirus early transcriptional activator immediate early-0 (ie-0) and 2 (ie-2) transgenic silkworm line. We hybridized it with Cas9 transgenic line to produce a double-positive transgenic Cas9(+)/sgIE0-sgIE2(+) line that could activate the CRISPR gene editing system. We first demonstrated that the system is capable of efficiently editing target genes and resulting in fragment deletions in the BmNPV genome. Survival rate of the transgenic Cas9(+)/sgIE0-sgIE2(+) line reached 65% after inoculation with 1 × 10 occlusion bodies/larva. Molecular analysis showed that BmNPV DNA replication and viral gene expression level in the transgenic Cas9(+)/sgIE0-sgIE2(+) line were significantly inhibited compared with the control Cas9(-)/sgIE0-sgIE2(-) line. These results indicated that IE-0 and IE-2, as baculovirus early transcriptional activators, can be used as target sites for gene therapy and that multigene editing could expand the range of target sites for research to create silkworm resistance breeds.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

Dong

et al. 2018

Insect Molecular Biology

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“…We detected mRNA level of BmC‐LZM by RT‐PCR and we used the silkworm translation initiation factor 4A ( sw22934 ) gene as an internal control. To examine viral DNA replication, we performed qPCR as previously described (Dong et al ., , ; Zhang et al ., ). The gp41 , viral gene, was used to quantify viral DNA load and we performed absolute quantification with a standard to detect gp41 copies.…”

Section: Methodsmentioning

confidence: 97%

Specific genes related to nucleopolyhedrovirus in Bombyx mori susceptible and near‐isogenic resistant strains through comparative transcriptome analysis

Chen

Tan

et al. 2019

Insect Molecular Biology

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Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the primary pathogens that causes severe economic losses to sericulture. Comparative transcriptomics analysis has been widely applied to explore the antiviral mechanism in resistant strains. Here, to identify genes involved in BmNPV infection, we identified differentially expressed genes (DEGs) and performed weighted gene co‐expression network analysis (WGCNA) between two Bombyx mori strains: strain 871 (susceptible to BmNPV infection) and the near‐isogenic strain 871C (resistant to BmNPV). Our results showed that 400 genes were associated with resistance in strain 871C, and 76 genes were related to susceptibility in strain 871. In addition, the correlation analysis of DEGs and WGCNA showed that 40 genes related to resistance were highly expressed in the resistant strain. Among them, gene BGIBMGA004291 was the most noticeable. We further identified the effect of gene BGIBMGA004291, which encoded a multiprotein bridge factor 2 (MBF2) family member (MBF2‐10), on viral infection in cells. Our data suggested that MBF2‐10 inhibited viral infection. Taken together, this study showed specific module trait correlations related to viral infection in strains 871 and 871C, and we identified a resistance‐related gene. These findings suggested promising candidate genes with antiviral activity, aiding in the analysis of the antiviral molecular mechanisms in resistant strains.

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“…During AcMNPV infection, most miRNAs from the host are down-regulated (Mehrabadi et al, 2013), although exceptions exist, such as bantam , which is upregulated and negatively regulates viral gene expression and replication (Shi et al, 2016). Finally, it was shown that the miRNA pathway could be harnessed to control BmNPV infection using expression constructs of artificial miRNA precursors (Zhang J. et al, 2014). …”

Section: Types Of Virus Infections In Insects and The Interaction Witmentioning

confidence: 99%

Viral Delivery of dsRNA for Control of Insect Agricultural Pests and Vectors of Human Disease: Prospects and Challenges

et al. 2017

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RNAi is applied as a new and safe method for pest control in agriculture but efficiency and specificity of delivery of dsRNA trigger remains a critical issue. Various agents have been proposed to augment dsRNA delivery, such as engineered micro-organisms and synthetic nanoparticles, but the use of viruses has received relatively little attention. Here we present a critical view of the potential of the use of recombinant viruses for efficient and specific delivery of dsRNA. First of all, it requires the availability of plasmid-based reverse genetics systems for virus production, of which an overview is presented. For RNA viruses, their application seems to be straightforward since dsRNA is produced as an intermediate molecule during viral replication, but DNA viruses also have potential through the production of RNA hairpins after transcription. However, application of recombinant virus for dsRNA delivery may not be straightforward in many cases, since viruses can encode RNAi suppressors, and virus-induced silencing effects can be determined by the properties of the encoded RNAi suppressor. An alternative is virus-like particles that retain the efficiency and specificity determinants of natural virions but have encapsidated non-replicating RNA. Finally, the use of viruses raises important safety issues which need to be addressed before application can proceed.

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Inhibition of BmNPV replication in silkworm cells using inducible and regulated artificial microRNA precursors targeting the essential viral gene lef-11

Cited by 44 publications

References 35 publications

CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

CRISPR/Cas9‐mediated disruption of the immediate early‐0 and 2 as a therapeutic approach to Bombyx mori nucleopolyhedrovirus in transgenic silkworm

Specific genes related to nucleopolyhedrovirus in Bombyx mori susceptible and near‐isogenic resistant strains through comparative transcriptome analysis

Viral Delivery of dsRNA for Control of Insect Agricultural Pests and Vectors of Human Disease: Prospects and Challenges

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