Global Analysis of Baculovirus Autographa californica Multiple Nucleopolyhedrovirus Gene Expression in the Midgut of the Lepidopteran Host Trichoplusia ni

Shrestha, Anita; Bao, Kan; Chen, Yun‐Ru; Chen, Wenbo; Wang, Ping; Fei, Zhangjun; Blissard, Gary W.

doi:10.1128/jvi.01277-18

Cited by 28 publications

(46 citation statements)

References 82 publications

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“…NC_001623 and BioProject accession no. PRJNA484772) (14). The percentage of total reads mapping to T. ni genes declined gradually as the infection proceeded ( Fig.…”

Section: Resultsmentioning

confidence: 97%

“…Differential expression analysis of T. ni transcripts following AcMNPV infection. To analyze the midgut cell responses to AcMNPV infection, developmentally synchronized and newly molted 5th-instar T. ni larvae were orally infected with wild-type (WT) AcMNPV OBs as described previously (14). Each larva was orally inoculated with 7 ϫ 10 4 OBs, a dose determined empirically to be the minimum approximate dose resulting in maximal numbers of infected midgut cells.…”

Section: Resultsmentioning

confidence: 99%

“…regulation of biosynthetic process (10) carbohydrate metabolic process (10) regulation of nitrogen compound metabolic process (11) cellular component assembly (11) lipid metabolic process (12) regulation of cellular metabolic process (12) regulation of macromolecule metabolic process (12) regulation of primary metabolic process (12) single-organism biosynthetic process (13) cellular response to stress (14) cellular developmental process (14) phosphorus metabolic process (18) small molecule metabolic process (19) organic substance biosynthetic process (25) cellular biosynthetic process (25) organelle organization (29) transport (29) cell cycle (29) protein metabolic process (42) nucleobase-containing compound metabolic process (71) cellular aromatic compound metabolic process (73) macromolecule metabolic process (102) cellular macromolecule metabolic process (101) cellular nitrogen compound metabolic process (92) organic cyclic compound metabolic process (75) heterocycle metabolic process (73) Sequence Distribution [Biological Process] reproduction (10) homeostatic process (11) intracellular transport (11) multicellular organismal process (12) growth (12) ribonucleoprotein complex assembly (13) mitotic cell cycle (13) nucleobase-containing compound catabolic process (15) protein folding (15) carbohydrate metabolic process (15) cofactor metabolic process (19) translation (22) lipid metabolic process (23) tRNA metabolic process (23) cell differentiation (28) mRNA processing (30) single-organism biosynthetic process (34) signal transduction (37) cellular protein modification process…”

Section: Suppressed Transcripts C Induced Transcripts Graph Levelmentioning

confidence: 99%

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Transcriptional Responses of the Trichoplusia ni Midgut to Oral Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus

Shrestha

Bao

Chen

et al. 2019

J Virol

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Baculoviruses are large double-stranded DNA viruses that are virulent pathogens of certain insect species. In a natural host, Trichoplusia ni, infection by the model baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) begins when the occluded form of the virus disassembles in the midgut and virions infect midgut epithelial cells to establish the primary phase of the infection. To better understand the primary phase of the AcMNPV infection cycle, newly molted 5th-instar T. ni larvae were orally infected with AcMNPV occlusion bodies and the transcriptional responses of the T. ni midgut were analyzed at various times from 0 to 72 h postinfection, using transcriptome sequencing analysis and a T. ni reference genome. The numbers of differentially expressed host genes increased as the infection progressed, and we identified a total of 3,372 differentially expressed T. ni transcripts in the AcMNPV-infected midgut. Genes encoding orthologs of HMG176, atlastin, and CPH43 were among the most dramatically upregulated in response to AcMNPV infection. A number of cytochrome P450 genes were downregulated in response to infection. We also identified the effects of AcMNPV infection on a large variety of genes associated with innate immunity. This analysis provides an abundance of new and detailed information on host responses to baculovirus infection during the primary phase of the infection in the midgut and will be important for understanding how baculoviruses establish productive infections in the organism. IMPORTANCE Baculoviruses are virulent pathogens of a number of important insect pest species. In the host Trichoplusia ni, infection begins in the midgut when infectious virions of the occlusion-derived virus (ODV) phenotype enter and subsequently replicate in cells of the midgut epithelium. A second virion phenotype (budded virus [BV]) is produced there, and BV mediates systemic infection of the animal. Most prior detailed studies of baculovirus infections have focused on BV infections of cultured cells. In this study, we examined the transcriptional responses of the T. ni midgut to infection by ODV of the baculovirus AcMNPV and identified a variety of host genes that respond dramatically to viral infection. Understanding the transcriptional responses of the host midgut to viral infection is critically important for understanding the biphasic infection in the animal as a whole.

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“…NC_001623 and BioProject accession no. PRJNA484772) (14). The percentage of total reads mapping to T. ni genes declined gradually as the infection proceeded ( Fig.…”

Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Suppressed Transcripts C Induced Transcripts Graph Levelmentioning

confidence: 99%

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Transcriptional Responses of the Trichoplusia ni Midgut to Oral Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus

Shrestha

Bao

Chen

et al. 2019

J Virol

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“…They are widely used as biopesticide as well as versatile and powerful vector for recombinant protein expression (Airenne et al, ). In nature, baculoviruses mainly infect Lepidoptera larvae when they feed on plant contaminated with the virus (Shrestha et al, ). During their viral biphasic life cycle, baculoviruses produce two distinct virion phenotypes: Occlusion‐derived viruses (ODVs) and budded viruses (BVs).…”

Section: Introductionmentioning

confidence: 99%

Improvement of baculovirus as protein expression vector and as biopesticide by CRISPR/Cas9 editing

Pazmiño-Ibarra

Mengual-Martí

Targovnik

et al. 2019

Biotech & Bioengineering

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The clustered regularly interspaced short palindromic repeats (CRISPR) system–associated Cas9 endonuclease is a molecular tool that enables specific sequence editing with high efficiency. In this study, we have explored the use of CRISPR/Cas9 system for the engineering of baculovirus. We have shown that the delivering of Cas9‐single guide RNA ribonucleoprotein (RNP) complex with or without DNA repair template into Sf21 insect cells through lipofection might be efficient to produce knockouts as well as knock‐ins into the baculovirus. To evaluate potential application of our CRISPR/Cas9 method to improve baculovirus as protein expression vector and as biopesticide, we attempted to knockout several genes from a recombinant AcMNPV form used in the baculovirus expression system as well as in a natural occurring viral isolate from the same virus. We have additionally confirmed the adaptation of this methodology for the generation of viral knock‐ins in specific regions of the viral genome. Analysis of the generated mutants revealed that the editing efficiency and the type of changes was variable but relatively high. Depending on the targeted gene, the editing rate ranged from 10% to 40%. This study established the first report revealing the potential of CRISPR/Cas9 for genome editing in baculovirus, contributing to the engineering of baculovirus as a protein expression vector as well as a biological control agent.

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“…They are widely used as biopesticide as well as versatile and powerful vector for recombinant protein expression (2). In nature, the baculovirus mainly infects Lepidoptera larvae when they feed on plant contaminated with the virus (3). During their viral biphasic life cycle, baculoviruses produce two distinct virion phenotypes: occlusion derived viruses (ODV) and budded viruses (BV).…”

Section: Introductionmentioning

confidence: 99%

Improvement of baculovirus as protein expression vector and as biopesticide by CRISPR/Cas9 editing

Pazmiño-Ibarra

Mengual

Targovnik

et al. 2019

Preprint

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adaptation of this methodology for the generation of viral knocks-in specific regions of the viral 35 genome. Analysis of the generated mutants revealed that the edition efficiency and the type of 36 changes was variable but relatively high. Depending on the targeted gene, the rate of edition 37 ranged from 10% to 40%. This study established the first report revealing the potential of 38 CRISPR/Cas9 for the edition of baculovirus contributing to the engineering of baculovirus as 39 protein expression vector as well as a biological control agent. 40 Keywords: CRISPR/Cas9, baculovirus, AcMNPV, genome editing, knock-out, knock-in 41 42 43 with the virus (3). During their viral biphasic life cycle, baculoviruses produce two distinct virion 49 phenotypes: occlusion derived viruses (ODV) and budded viruses (BV). While ODV are 50 involved in horizontal virus transmission from insect to insect through structures named 51 occlusion bodies (OB) which have the virus embedded within, BV are involved in spread of the 52 infection from cell to cell (4). 53Autographa californica multiple nucleopolyhedrovirus (AcMNPV), the prototypic baculovirus 54 most common used for biotechnological purpose, has a circular genome of about 134 Kb that 55contains 156 predicted open reading frames (ORF) (5). The baculovirus genomes, and specially 56 AcMNPV genome, have been genetically modified in order to enhance their pesticide potency 57 and increase the quality and quantity of the recombinant protein expressed in the system (6, 7). 58These modifications include either the deletion of non-essential genes for virus survival or 59 infectivity and the insertion of foreign genes. For instance, in term of increase the insecticidal 60 activity of baculovirus, the cry1Ab gene from Bacillus thuringensis and neurotoxins from 61 scorpion venom have been incorporated into the baculovirus genome (8, 9). In addition, the egt 62 gene (Ac15), involved in the inhibition of the molting, has been deleted from the virus 63 significantly improving the speed of killing of the virus (10). Moreover, different attempt has 64 been conducted to improve the heterologous protein expression in the baculovirus expression 65 system (BEVS). It has been reported that simultaneous deletion of non-essential genes for the in 66 produce insertions or deletions (indels), or homology-directed repair (HDR) in presence of a 87 donor template DNA with homology to the sequence flanking the DSB (16-18). Recent 88 developments in this technology have made possible to generated precise modifications into a 89 5 wide variety of viral genomes (19). However, the CRISPR/Cas9 system has not been applied for 90 the edition of baculoviral genomes. 91In this study, we developed for first time a CRISPR/Cas9-assisted method to edit AcMNPV 92 genome in multiple ways and with different purposes. We have shown that the delivering of 93 Cas9-sgRNA ribonucleoprotein (RNP) complex through lipofection in insect cells might be 94 efficient to generate gene knock-out and knock-in. To evaluate potential app...

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Global Analysis of Baculovirus Autographa californica Multiple Nucleopolyhedrovirus Gene Expression in the Midgut of the Lepidopteran Host Trichoplusia ni

Cited by 28 publications

References 82 publications

Transcriptional Responses of the Trichoplusia ni Midgut to Oral Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus

Transcriptional Responses of the Trichoplusia ni Midgut to Oral Infection by the Baculovirus Autographa californica Multiple Nucleopolyhedrovirus

Improvement of baculovirus as protein expression vector and as biopesticide by CRISPR/Cas9 editing

Improvement of baculovirus as protein expression vector and as biopesticide by CRISPR/Cas9 editing

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