Publication: Large-scale production and antiviral efficacy of multi-target double-stranded RNA for the prevention of white spot syndrome virus (WSSV) in shrimp
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Issued Date
2015
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eng
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Mahidol University
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BioMed Central
Bibliographic Citation
BMC Biotechnology. Vol. 15, (2015), 110
Suggested Citation
Thitiporn Thammasorn, Pakkakul Sangsuriya, Watcharachai Meemetta, Saengchan Senapin, Sarocha Jitrakorn, Triwit Rattanarojpong, Vanvimon Saksmerprome Large-scale production and antiviral efficacy of multi-target double-stranded RNA for the prevention of white spot syndrome virus (WSSV) in shrimp. BMC Biotechnology. Vol. 15, (2015), 110. doi:DOI 10.1186/s12896-015-0226-9 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/2737
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Title
Large-scale production and antiviral efficacy of multi-target double-stranded RNA for the prevention of white spot syndrome virus (WSSV) in shrimp
Abstract
Background: RNA interference (RNAi) is a specific and effective approach for inhibiting viral replication by
introducing double-stranded (ds)RNA targeting the viral gene. In this study, we employed a combinatorial
approach to interfere multiple gene functions of white spot syndrome virus (WSSV), the most lethal shrimp
virus, using a single-batch of dsRNA, so-called “multi-WSSV dsRNA.” A co-cultivation of RNase-deficient E. coli
was developed to produce dsRNA targeting a major structural protein (VP28) and a hub protein (WSSV051)
with high number of interacting protein partners.
Results: For a co-cultivation of transformed E. coli, use of Terrific broth (TB) medium was shown to improve
the growth of the E. coli and multi-WSSV dsRNA yields as compared to the use of Luria Bertani (LB) broth. Co-culture
expression was conducted under glycerol feeding fed-batch fermentation. Estimated yield of multi-WSSV dsRNA
(μg/mL culture) from the fed-batch process was 30 times higher than that obtained under a lab-scale culture with LB
broth. Oral delivery of the resulting multi-WSSV dsRNA reduced % cumulative mortality and delayed average time to
death compared to the non-treated group after WSSV challenge.
Conclusion: The present study suggests a co-cultivation technique for production of antiviral dsRNA with
multiple viral targets. The optimal multi-WSSV dsRNA production was achieved by the use of glycerol feeding
fed-batch cultivation with controlled pH and dissolved oxygen. The cultivation technique developed herein
should be feasible for industrial-scale RNAi applications in shrimp aquaculture. Interference of multiple viral
protein functions by a single-batch dsRNA should also be an ideal approach for RNAi-mediated fighting
against viruses, especially the large and complicated WSSV.