The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice

Feng, Zhenqing; Chen, Jianghua; Liang, Wangwang; Chen, Wenzhi; Li, Zhaolong; Chen, Qi; Cai, Shaoli

doi:10.1186/s12985-020-01450-7

Cited by 29 publications

(31 citation statements)

References 54 publications

(82 reference statements)

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“…PRV is a model organism for studying the molecular pathogenesis of herpesviruses, including the mechanism of neurotropism [6,7] and the regulation of gene expression. PRV also serves as a model for the development of genetically engineered vaccines [8].…”

Section: Introductionmentioning

confidence: 99%

An Integrated Sequencing Approach for Updating the Pseudorabies Virus Transcriptome

et al. 2021

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In the last couple of years, the implementation of long-read sequencing (LRS) technologies for transcriptome profiling has uncovered an extreme complexity of viral gene expression. In this study, we carried out a systematic analysis on the pseudorabies virus transcriptome by combining our current data obtained by using Pacific Biosciences Sequel and Oxford Nanopore Technologies MinION sequencing with our earlier data generated by other LRS and short-read sequencing techniques. As a result, we identified a number of novel genes, transcripts, and transcript isoforms, including splice and length variants, and also confirmed earlier annotated RNA molecules. One of the major findings of this study is the discovery of a large number of 5′-truncations of larger putative mRNAs being 3′-co-terminal with canonical mRNAs of PRV. A large fraction of these putative RNAs contain in-frame ATGs, which could initiate translation of N-terminally truncated polypeptides. Our analyses indicate that CTO-S, a replication origin-associated RNA molecule is expressed at an extremely high level. This study demonstrates that the PRV transcriptome is much more complex than previously appreciated.

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Section: Introductionmentioning

confidence: 99%

An Integrated Sequencing Approach for Updating the Pseudorabies Virus Transcriptome

et al. 2021

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“…PRV is a model organism for studying the molecular pathogenesis of herpesviruses, including the mechanism of neurotropism [6,7] and the regulation of gene expression. PRV is also served as a model for the development of genetically engineered vaccines [8]. Additionally, this virus is the most popular multisynaptic tracer of Direct RNA sequencing generates less false transcripts, but it produces incomplete reads since 15-30 bp sequences always lack from the 5'-termini, and in many cases poly(A) tails are also missing.…”

Section: Introductionmentioning

confidence: 99%

An Integrated Sequencing Approach for Updating of Pseudorabies Virus Transcriptome

Torma¹,

Tombácz²,

Csabai³

et al. 2020

Preprint

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In the last couple of years, the implementation of long-read sequencing (LRS) technologies for transcriptome profiling has uncovered an extreme complexity of viral gene expression. In this study, we carried out a systematic analysis on the pseudorabies virus transcriptome by combining our current data obtained by using Pacific Biosciences Sequel and Oxford Nanopore Technologies MinION sequencings with our earlier data generated by other LRS and short-read sequencing techniques. As a result, we identified a number of novel genes, transcripts, and transcript isoforms, including splice and length variants, and also confirmed earlier annotated RNA molecules. One of the major findings of this study is the discovery of a large number of 5’-truncated putative mRNAs embedded into larger host mRNAs. A large fraction of these RNA molecules contain in-frame ORFs, which may encode N-terminally truncated polypeptides. These study demonstrates that the PRV transcriptome is much more complex than previously appreciated.

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“…However, due to the different situations in vitro and in vivo, it is worthwhile to detect and compare the expression of the three recombinant viruses in vivo in further studies. A previous study has shown that the viral genome DNA was detected in the brain, heart, lung, liver, kidney, and spleen tissues of PRV-Fa infected mice, but not in PRV-ΔgE/ΔgI/ΔTK infected group at 72 h post infection [ 6 ]. When the infection time was extended to 200 h, viral DNA was detected in the brain and lungs of mice infected with PRV-ΔgE/ΔgI/ΔTK.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, we generated a TK&gE-deleted PRV (PRV Δ TK&gE−AH02 ), which is safe for 1-day-old piglets with maternal PRV antibodies and 4 ~ 5 week-old PRV antibody negative piglets and can provide complete protection in weaned pigs against challenged with virulent AH02LA strain at 7 days post vaccination [ 4 ]. Several previous reports have shown that recombinant PRVs expressing foreign antigens are able to stimulate humoral and cell-mediated immune responses against both PRV and other viruses [ 5 , 6 ]. Thus, PRV Δ TK&gE−AH02 might be a technically appropriate vector for the expression of foreign antigens of other swine diseases.…”

Section: Introductionmentioning

confidence: 99%

Identification of four insertion sites for foreign genes in a pseudorabies virus vector

Zhang

Guo

et al. 2021

BMC Vet Res

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Background Pseudorabies virus (PRV) is a preferred vector for recombinant vaccine construction. Previously, we generated a TK&gE-deleted PRV (PRVΔTK&gE−AH02) based on a virulent PRV AH02LA strain. It was shown to be safe for 1-day-old piglets with maternal PRV antibodies and 4 ~ 5 week-old PRV antibody negative piglets and provide rapid and 100 % protection in weaned pigs against lethal challenge with the PRV variant strain. It suggests that PRVTK&gE−AH02 may be a promising live vaccine vector for construction of recombinant vaccine in pigs. However, insertion site, as a main factor, may affect foreign gene expression. Results In this study, we constructed four recombinant PRV-S bacterial artificial chromosomes (BACs) carrying the same spike (S) expression cassette of a variant porcine epidemic diarrhea virus strain in different noncoding regions (UL11-10, UL35-36, UL46-27 or US2-1) from AH02LA BAC with TK, gE and gI deletion. The successful expression of S gene (UL11-10, UL35-36 and UL46-27) in recombinant viruses was confirmed by virus rescue, PCR, real-time PCR and indirect immunofluorescence. We observed higher S gene mRNA expression level in swine testicular cells infected with PRV-S(UL11-10)ΔTK/gE and PRV-S(UL35-36)ΔTK/gE compared to that of PRV-S(UL46-27)ΔTK/gE at 6 h post infection (P < 0.05). Moreover, at 12 h post infection, cells infected with PRV-S(UL11-10)ΔTK/gE exhibited higher S gene mRNA expression than those infected with PRV-S(UL35-36)ΔTK/gE (P = 0.097) and PRV-S(UL46-27)ΔTK/gE (P < 0.05). Recovered vectored mutant PRV-S (UL11-10, UL35-36 and UL46-27) exhibited similar growth kinetics to the parental virus (PRVΔTK&gE−AH02). Conclusions This study focuses on identification of suitable sites for insertion of foreign genes in PRV genome, which laids a foundation for future development of recombinant PRV vaccines.

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The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice

Cited by 29 publications

References 54 publications

An Integrated Sequencing Approach for Updating the Pseudorabies Virus Transcriptome

An Integrated Sequencing Approach for Updating the Pseudorabies Virus Transcriptome

An Integrated Sequencing Approach for Updating of Pseudorabies Virus Transcriptome

Identification of four insertion sites for foreign genes in a pseudorabies virus vector

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