Using double-stranded RNA for the control of Laem-Singh Virus (LSNV) in Thai P. monodon

Saksmerprome, Vanvimon; Thammasorn, Thitiporn; Jitrakorn, Sarocha; Wongtripop, Somjai; Borwornpinyo, Suparerk; Withyachumnarnkul, Boonsirm

doi:10.1016/j.jbiotec.2013.01.028

Cited by 33 publications

(24 citation statements)

References 15 publications

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“…In the past decade, RNAi-based technology has gained attention for its potential to control shrimp viral diseases that have caused significant losses in aquaculture. Inhibition of shrimp viruses by viral-specific dsRNA were demonstrated in insect cell lines (He et al, 2009;Theerawanitchpan et al, 2012) and in penaeid shrimp (Robalino et al, 2005;Yodmuang et al, 2006;Ongvarrasopone et al, 2008;Saksmerprome et al, 2009Saksmerprome et al, , 2013Thammasorn et al, 2013). Given that sequences of viral genes of interest are available, dsRNA targeting those genes can be synthesized by in vitro transcription and in vivo bacterial system.…”

Section: Introductionmentioning

confidence: 99%

Use of microalgae Chlamydomonas reinhardtii for production of double-stranded RNA against shrimp virus

Somchai

Jitrakorn

Thitamadee

et al. 2016

Aquaculture Reports

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a b s t r a c t RNA interference has been proposed to be a promising tool for combating shrimp viruses. Antiviral double-stranded (ds)RNA has been mostly produced in Escherichia coli-expression system because of its high efficiency and inexpensive operations. However, overusing the bacteria may raise concerns regarding public health and environmental contamination, and seeking for a new dsRNA production platform would be alternative for future molecular farming. In this study, we exploited the green microalgae Chlamydomonas reinhardtii to produce dsRNA targeting the lethal shrimp yellow head virus (YHV). The expression plasmid pSL18 for C. reinhardtii was constructed to contain YHV-specific hairpin RNA expression cassette, and the successful assembly of pSL18-YHV was confirmed by PCR and enzymatic digestions. Glass bead method was employed for transformation of C. reinhardtii nuclear genome with pSL18-YHV. Microalgal expression of dsRNA-YHV, approximately 45 ng from 100-mL culture, was detected by qRT-PCR. Oral feeding experiment on postlarval shrimp revealed that the formulated feed with C. reinhardtii expressing dsRNA-YHV, at the ratio of 1 × 10 8 transformants per gram feed, improved 22% survival rate after YHV challenge. The present study suggests that C. reinhardtii can be bioengineered to produce viralspecific dsRNA for shrimp viral disease control, and the developed qRT-PCR could detect microalgal dsRNA with detection limit of subpicogram.

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Section: Introductionmentioning

confidence: 99%

Use of microalgae Chlamydomonas reinhardtii for production of double-stranded RNA against shrimp virus

Somchai

Jitrakorn

Thitamadee

et al. 2016

Aquaculture Reports

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“…Transformed E. coli expressing dsRNA-LSNV was prepared according to the method described by Saksmerprome et al (2013).…”

Section: Preparation Of Transformed E Coli Variantsmentioning

confidence: 99%

“…In addition, the Artemia enrichment process was carried out using E. coli expressing dsRNA-LSNV that is previously shown to inhibit LSNV by injection and oral delivery (Saksmerprome et al, 2013). It is important that the dsLSNV-enrichment period was carried out soon after Artemia hatched or became Instar I nauplii, so Artemia remained the most nutritious diet for shrimp postlarvae.…”

Section: Successful Artemia Enrichment With E Coli Expressing Rfp Anmentioning

confidence: 99%

“…However, most of the successful reports are the results of laboratory efforts. A limited number of work on improving ''drug'' delivery methods for shrimp farming include applications of chitosan nanoparticles and the recombinant bacteria strains (Sarathi et al, 2008;Theerawanitchpan et al, 2012;Attasart et al, 2013;Saksmerprome et al, 2013). Therefore, it is much needed to seek the efficient and inexpensive methods to deliver therapeutics/protective agents for farm applications.…”

Section: Introductionmentioning

confidence: 99%

“…Retarded growth and wide variation of size (>35% of coefficient of variation) are among the main characteristics of MSGS shrimp that leads to economical loss in Thai shrimp farms since 2002. Here, Escherichia coli (E. coli)-expressed dsRNA specific to LSNV previously shown to effectively inhibit the virus by injection and oral administration (Saksmerprome et al, 2013) was encapsulated in Artemia via the enrichment process. P. monodon postlarvae (PL1-15) were fed with the enriched Artemia nauplii, and were examined for the antiviral effect against LSNV in comparison to the control group.…”

Section: Introductionmentioning

confidence: 99%

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