Comparison of factors that may affect the inhibitory efficacy of transgenic RNAi targeting of baculoviral genes in silkworm, Bombyx mori

Jiang, Liang; Zhao, Ping; Wang, Genhong; Cheng, Tingcai; Yang, Qiong; Jin, Shengkai; Lin, Ping; Xiao, Yang; Sun, Qiang; Xia, Qingyou

doi:10.1016/j.antiviral.2012.12.020

Cited by 55 publications

(33 citation statements)

References 67 publications

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“…In contrast, TG RNAi has been applied extensively by silencing one or multiple essential BmNPV genes to destroy specific viral mRNAs (7, 8). It is important to select appropriate genes as targets in RNAi-based anti-BmNPV experiments, as silencing of different BmNPV genes results in variable antiviral resistance efficiency (9). Overall, moderate virus resistance has been reported for TG RNAi-based anti-BmNPV strategies, thus leaving the major challenge of how to inhibit viral DNA replication due to the persistence of BmNPV genomic DNA.…”

Section: Introductionmentioning

confidence: 99%

Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus

Chen

Hou

et al. 2017

J Virol

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We developed a novel antiviral strategy by combining transposon-based transgenesis and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system for the direct cleavage of Bombyx mori nucleopolyhedrovirus (BmNPV) genome DNA to promote virus clearance in silkworms. We demonstrate that transgenic silkworms constitutively expressing Cas9 and guide RNAs targeting the BmNPV immediate early-1 (ie-1) and me53 genes effectively induce target-specific cleavage and subsequent mutagenesis, especially large (∼7-kbp) segment deletions in BmNPV genomes, and thus exhibit robust suppression of BmNPV proliferation. Transgenic animals exhibited higher and inheritable resistance to BmNPV infection than wild-type animals. Our approach will not only contribute to modern sericulture but also shed light on future antiviral therapy.IMPORTANCE Pathogen genome targeting has shown its potential in antiviral research. However, transgenic CRISPR/Cas9 system-mediated viral genome targeting has not been reported as an antiviral strategy in a natural animal host of a virus. Our data provide an effective approach against BmNPV infection in a real-world biological system and demonstrate the potential of transgenic CRISPR/Cas9 systems in antiviral research in other species.

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Section: Introductionmentioning

confidence: 99%

Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus

Chen

Hou

et al. 2017

J Virol

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“…Serious pathogens such as B. mori cytoplasmic polyhedrosis virus (BmCPV) and B. mori densovirus (BmDNV) infect only the silkworm midgut (Jiang and Xia, 2014). Silencing viral genes could enhance host resistance (Jiang and Xia, 2014;Jiang et al, 2013c). We are constructing transgenic silkworms with RNAi against multiple BmCPV and BmDNV genes using P3Pþ5UI.…”

Section: Discussionmentioning

confidence: 99%

“…Embryo microinjection results are in Supplementary Table 2. A single transgenic line was selected at random for each vector; lines were named P2þ5UI and P3þ5UI. Insertion sites of transgenic silkworms were analyzed as previously described (Jiang et al, , 2013c.…”

Section: Construction Of Transgenic Silkwormsmentioning

confidence: 99%

The 5′-UTR intron of the midgut-specific BmAPN4 gene affects the level and location of expression in transgenic silkworms

Jiang

Huang

Sun

et al. 2015

Insect Biochemistry and Molecular Biology

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“…The glycoprotein gene plays a key role in the progress that baculovirus infects cells and progeny nuclear capsid effective budding release [26, 27]. Besides, silencing glycoprotein gene in transgenic silkworm increases resistance to BmNPV [28, 29]. Therefore, the glycoprotein is one of the most important capsule membrane proteins in baculovirus.…”

Section: Introductionmentioning

confidence: 99%

Analysis of codon usage bias of envelope glycoprotein genes in nuclear polyhedrosis virus (NPV) and its relation to evolution

Zhao

Zheng

et al. 2016

BMC Genomics

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BackgroundAnalysis of codon usage bias is an extremely versatile method using in furthering understanding of the genetic and evolutionary paths of species. Codon usage bias of envelope glycoprotein genes in nuclear polyhedrosis virus (NPV) has remained largely unexplored at present. Hence, the codon usage bias of NPV envelope glycoprotein was analyzed here to reveal the genetic and evolutionary relationships between different viral species in baculovirus genus.ResultsA total of 9236 codons from 18 different species of NPV of the baculovirus genera were used to perform this analysis. Glycoprotein of NPV exhibits weaker codon usage bias. Neutrality plot analysis and correlation analysis of effective number of codons (ENC) values indicate that natural selection is the main factor influencing codon usage bias, and that the impact of mutation pressure is relatively smaller. Another cluster analysis shows that the kinship or evolutionary relationships of these viral species can be divided into two broad categories despite all of these 18 species are from the same baculovirus genus.ConclusionsThere are many elements that can affect codon bias, such as the composition of amino acids, mutation pressure, natural selection, gene expression level, and etc. In the meantime, cluster analysis also illustrates that codon usage bias of virus envelope glycoprotein can serve as an effective means of evolutionary classification in baculovirus genus.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3021-7) contains supplementary material, which is available to authorized users.

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Comparison of factors that may affect the inhibitory efficacy of transgenic RNAi targeting of baculoviral genes in silkworm, Bombyx mori

Cited by 55 publications

References 67 publications

Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus

Transgenic Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-Mediated Viral Gene Targeting for Antiviral Therapy of Bombyx mori Nucleopolyhedrovirus

The 5′-UTR intron of the midgut-specific BmAPN4 gene affects the level and location of expression in transgenic silkworms

Analysis of codon usage bias of envelope glycoprotein genes in nuclear polyhedrosis virus (NPV) and its relation to evolution

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