Cargando…

Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain

BACKGROUND: Viruses cause significant economic losses to shrimp aquaculture worldwide. In severe cases, they can lead to 100% mortality within a matter of days, hence the aquaculture industry requires antiviral strategies to minimize economic impacts. Currently, a double-stranded RNA (dsRNA)-based p...

Descripción completa

Detalles Bibliográficos
Autores principales: Wuthisathid, Kitti, Chaijarasphong, Thawatchai, Chotwiwatthanakun, Charoonroj, Somrit, Monsicha, Sritunyalucksana, Kallaya, Itsathitphaisarn, Ornchuma
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7989029/
https://www.ncbi.nlm.nih.gov/pubmed/33757419
http://dx.doi.org/10.1186/s12866-021-02148-8
_version_ 1783668881193172992
author Wuthisathid, Kitti
Chaijarasphong, Thawatchai
Chotwiwatthanakun, Charoonroj
Somrit, Monsicha
Sritunyalucksana, Kallaya
Itsathitphaisarn, Ornchuma
author_facet Wuthisathid, Kitti
Chaijarasphong, Thawatchai
Chotwiwatthanakun, Charoonroj
Somrit, Monsicha
Sritunyalucksana, Kallaya
Itsathitphaisarn, Ornchuma
author_sort Wuthisathid, Kitti
collection PubMed
description BACKGROUND: Viruses cause significant economic losses to shrimp aquaculture worldwide. In severe cases, they can lead to 100% mortality within a matter of days, hence the aquaculture industry requires antiviral strategies to minimize economic impacts. Currently, a double-stranded RNA (dsRNA)-based platform has been proven effective at a laboratory scale. The bottleneck for its industrialization is the lack of low-cost, efficient and practical delivery approaches. In an effort to bridge the gap between laboratory and farm applications, virus-like particles (VLP) have been used as nanocarriers of dsRNA. However, the implementation of this approach still suffers from high costs and a lengthy procedure, co-expression of subunits of VLP or capsid proteins (CPs) and dsRNA can be the solution for the problem. CP and dsRNA are traditionally expressed in two different E. coli hosts: protease-deficient and RNase III-deficient strains. To condense the manufacturing of dsRNA-containing VLP, this study constructed a novel E. coli strain that is able to co-express viral capsid proteins and dsRNA in the same E. coli cell. RESULTS: A novel bacterial strain DualX-B15(DE3) was engineered to be both protease- and RNase III-deficiency via P1 phage transduction. The results revealed that it could simultaneously express recombinant proteins and dsRNA. CONCLUSION: Co-expression of viral capsid proteins and dsRNA in the same cell has been shown to be feasible. Not only could this platform serve as a basis for future cost-effective and streamlined production of shrimp antiviral therapeutics, it may be applicable for other applications that requires co-expression of recombinant proteins and dsRNA. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02148-8.
format Online
Article
Text
id pubmed-7989029
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-79890292021-03-25 Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain Wuthisathid, Kitti Chaijarasphong, Thawatchai Chotwiwatthanakun, Charoonroj Somrit, Monsicha Sritunyalucksana, Kallaya Itsathitphaisarn, Ornchuma BMC Microbiol Research Article BACKGROUND: Viruses cause significant economic losses to shrimp aquaculture worldwide. In severe cases, they can lead to 100% mortality within a matter of days, hence the aquaculture industry requires antiviral strategies to minimize economic impacts. Currently, a double-stranded RNA (dsRNA)-based platform has been proven effective at a laboratory scale. The bottleneck for its industrialization is the lack of low-cost, efficient and practical delivery approaches. In an effort to bridge the gap between laboratory and farm applications, virus-like particles (VLP) have been used as nanocarriers of dsRNA. However, the implementation of this approach still suffers from high costs and a lengthy procedure, co-expression of subunits of VLP or capsid proteins (CPs) and dsRNA can be the solution for the problem. CP and dsRNA are traditionally expressed in two different E. coli hosts: protease-deficient and RNase III-deficient strains. To condense the manufacturing of dsRNA-containing VLP, this study constructed a novel E. coli strain that is able to co-express viral capsid proteins and dsRNA in the same E. coli cell. RESULTS: A novel bacterial strain DualX-B15(DE3) was engineered to be both protease- and RNase III-deficiency via P1 phage transduction. The results revealed that it could simultaneously express recombinant proteins and dsRNA. CONCLUSION: Co-expression of viral capsid proteins and dsRNA in the same cell has been shown to be feasible. Not only could this platform serve as a basis for future cost-effective and streamlined production of shrimp antiviral therapeutics, it may be applicable for other applications that requires co-expression of recombinant proteins and dsRNA. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02148-8. BioMed Central 2021-03-23 /pmc/articles/PMC7989029/ /pubmed/33757419 http://dx.doi.org/10.1186/s12866-021-02148-8 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Wuthisathid, Kitti
Chaijarasphong, Thawatchai
Chotwiwatthanakun, Charoonroj
Somrit, Monsicha
Sritunyalucksana, Kallaya
Itsathitphaisarn, Ornchuma
Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title_full Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title_fullStr Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title_full_unstemmed Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title_short Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain
title_sort co-expression of double-stranded rna and viral capsid protein in the novel engineered escherichia coli dualx-b15(de3) strain
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7989029/
https://www.ncbi.nlm.nih.gov/pubmed/33757419
http://dx.doi.org/10.1186/s12866-021-02148-8
work_keys_str_mv AT wuthisathidkitti coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain
AT chaijarasphongthawatchai coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain
AT chotwiwatthanakuncharoonroj coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain
AT somritmonsicha coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain
AT sritunyalucksanakallaya coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain
AT itsathitphaisarnornchuma coexpressionofdoublestrandedrnaandviralcapsidproteininthenovelengineeredescherichiacolidualxb15de3strain