DNA constructs expressing long-hairpin RNA (lhRNA) protect Penaeus monodon against White Spot Syndrome Virus

Krishnan, P.; Babu, P. Gireesh; Saravanan, S.; Rajendran, K. V.; Chaudhari, Aparna

doi:10.1016/j.vaccine.2009.04.011

Cited by 35 publications

(34 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The WSSV structural genes that have been targeted for silencing so far by RNAi technology include vp15 (Westenberg et al 2005), vp19 (Robalino et al 2004, 2005, Krishnan et al 2009), vp28 (Westenberg et al 2005, Kim et al 2007, Xu et al 2007, Krishnan et al 2009) and vp281 (Kim et al 2007). YHV genes that have been used as the targets for RNAi-based gene silencing trials are protease (Tirasophon et al 2005, 2007, RNA polymerase , helicase and RdRP (Tirasophon et al 2005).…”

Section: Target Genementioning

confidence: 99%

“…The extent of the threat posed by these suppressors to RNAi-based therapeutics is not fully established. However, a number of recent studies showing RNAi-induced viral gene silencing corroborate that RNAi-based viral therapies are potential tools for disease management (Novina et al 2002, Chen et al 2006a, Kusar et al 2006, Krishnan et al 2009). …”

mentioning

confidence: 98%

“…siRNAs may also be synthesized by in vitro digestion of bacterially expressed long dsRNA with the Dicer enzyme. Plasmid constructs that express lhRNA in vivo have been employed to silence the vp19 and vp28 genes of WSSV in tiger shrimp Penaeus monodon (Krishnan et al 2009). As discussed earlier, employing shRNA and lhRNA expression constructs for this purpose facilitates long-term silencing of the viral genes.…”

mentioning

confidence: 99%

See 2 more Smart Citations

RNA interference-based therapeutics for shrimp viral diseases

Krishnan¹,

Gireesh‐Babu²,

Rajendran³

et al. 2009

Dis. Aquat. Org.

View full text Add to dashboard Cite

RNA interference (RNAi) has emerged as a powerful tool to manipulate gene expression in the laboratory. The presence of a double-stranded RNA (dsRNA) in eukaryotic cells triggers this post-transcriptional gene-silencing mechanism, leading to a sequence-specific degradation of the target mRNA. Among its many potential biomedical applications, silencing of viral genes stands out as a promising therapeutic strategy. Marine shrimp viral diseases, especially white spot disease (WSD), represents one of the most attractive targets for the development of therapeutic RNAi owing to its widespread economic impact. This review summarizes the current knowledge in the therapeutic application of RNAi for combating viral diseases in shrimp. The basic principles of RNAi are described, focusing on features important for its therapeutic manipulation. Subsequently, a stepwise strategy for the development of therapeutic RNAi is presented.KEY WORDS: RNAi therapy · Shrimp diseases · Small interfering RNA · siRNA · Long hairpin RNA · lhRNA Resale or republication not permitted without written consent of the publisherDis Aquat Org 86: [263][264][265][266][267][268][269][270][271][272] 2009 exception of MSGS, are classified as OIE notifiable (OIE 2006). Among these, WSSV has had the greatest impact on shrimp farming and is currently considered the most important shrimp disease in terms of distribution and economic losses. Current therapeutic strategiesManaging viral diseases has been the greatest challenge to the shrimp industry because short-term strategies, such as the use of immunostimulants, bioremediators and probiotics, have their own limitations in terms of their efficacy, practicability, cost, and, above all, reproducibility. Unlike finfishes, the shrimps lack a developed adaptive immune system, which is a prerequisite for the development of any protein vaccines. Subunit vaccines (Witteveldt et al. 2004a(Witteveldt et al. ,b, 2006 have been reported to provide some degree of protection to shrimps from viral infection, but their field level potential is yet to be seen. In this scenario, RNAi, which seems to offer great promise in terms of treating human and animal diseases, is looked upon with great hope by shrimp health managers. ENDOGENOUS RNAi PATHWAYObservations were made in plants (Napoli et al. 1990, van der Krol et al. 1990) and nematodes (Lee et al. 1993),which hinted at the existence of the RNAi pathway years before double-stranded RNA (dsRNA) molecules were identified as the key component of this evolutionarily conserved post-transcriptional silencing pathway in eukaryotic cells (Fire et al. 1998). Since then, we have witnessed significant advances in the understanding of how RNAi functions and identified several effective ways to manipulate it in the laboratory. Although a detailed account on the mechanistic aspects of RNAi is beyond the scope of this review and can be found elsewhere (Bartel 2004, Carmell & Hannon 2004, Cullen 2004, Meister & Tuschl 2004, Murchison & Hannon 2004, Du & Zamore 2005, Kim 20...

show abstract

Section: Target Genementioning

confidence: 99%

mentioning

confidence: 98%

mentioning

confidence: 99%

See 1 more Smart Citation

RNA interference-based therapeutics for shrimp viral diseases

Krishnan¹,

Gireesh‐Babu²,

Rajendran³

et al. 2009

Dis. Aquat. Org.

View full text Add to dashboard Cite

show abstract

“…Lately, considerable scientific effort has been directed at exploring the use of RNAi molecules for controlling this devastating virus [8,10,15,28]. This strategy is particularly attractive because the rudimentary specific immune system of shrimp limits the success of conventional strategies like protein based vaccination [20,22,31,32] and use of immunostimulants [3].…”

Section: Introductionmentioning

confidence: 99%

“…This is also important in the light that despite being the most highly expressed protein, it was not reported as a major antigenic protein in numerous studies conducted since the virus was first reported in 1992. A recent study from our lab [10,11] demonstrated the efficacy of DNA constructs capable of expressing lhRNA against WSSV capsid genes (vp19 and vp28) in protecting Penaeus monodon from WSSV infection. The results showed VP28 to be more protective than VP19, which is why several researchers have targeted this protein for vaccine development [8,21,28,31].…”

Section: Introductionmentioning

confidence: 99%

Protection of Penaeus monodon from Infection of White spot syndrome virus by DNA Construct Expressing Long Hairpin-RNA Against ICP11 Gene

et al. 2010

Self Cite

View full text Add to dashboard Cite

A plasmid construct (pICP11-LH) was designed to constitutively express long-hairpin RNA (lhRNA) against icp11 gene, which is reportedly the most highly expressed gene of White spot syndrome virus (WSSV) and likely to have an important role in viral pathogenesis. The construct was used singly and in combination with other similar constructs designed against vp28 and vp19. A total of 6 treatments, T1 (pICP11-LH; 35 μg), T2 (pVP28-LH; 35 μg), T3 (pVP28-LH and pVP19-LH; 17.5 μg each), T4 (pVP28-LH and pVP19-LH; 25 μg:10 μg), T5 (pICP11-LH, pVP28-LH and pVP19-LH; 11.5 μg each) and T6 (pGFP-LH; 35 μg) were injected intramuscularly into 20 g Penaeus monodon specimens. The shrimp were challenged with WSSV 24 hpi and protection efficacy was measured in terms of survival and viral load 15 days after challenge. Appropriate negative and positive controls were used. T2 and T3 offered highest protection (75%) followed by T1 (67%) and T4 and T5 groups (58%), while T6 showed 25% protection. In all the target specific treatments, the viral load as estimated by single tube WSSV kit was kept in check (10-100 copies), whereas in the unimmunized challenged controls it progressed to severe infection (>10(5) copies). In spite of over 3 times higher expression of ICP11 compared to VP28, its knockdown by pICP11-LH did not offer any protective advantage over pVP28-LH, either singly or in combination. Moreover, none of the combinations bettered the protection efficacy of pVP28-LH administered alone. To investigate concerns about deleterious effect of plasmid persistence and constitutive expression on shrimp growth, a lab-scale 1 month growth study was conducted with 4 treatments T2, T3, T4 and T6, where no difference in specific growth rate was observed compared to controls.

show abstract

Design and Construction of Shrimp Antiviral DNA Vaccines Expressing Long and Short Hairpins for Protection by RNA Interference

2016

View full text Add to dashboard Cite

DNA vaccines present the aquaculture industry with an effective and economically viable method of controlling viral pathogens that drastically affect productivity. Since specific immune response is rudimentary in invertebrates, the presence of RNA interference (RNAi) pathway in shrimps provides a promising new approach to vaccination. Plasmid DNA vaccines that express short or long double stranded RNA in vivo have shown protection against viral diseases. The design, construction and considerations for preparing such vaccines are discussed.

show abstract

DNA constructs expressing long-hairpin RNA (lhRNA) protect Penaeus monodon against White Spot Syndrome Virus

Cited by 35 publications

References 48 publications

RNA interference-based therapeutics for shrimp viral diseases

RNA interference-based therapeutics for shrimp viral diseases

Protection of Penaeus monodon from Infection of White spot syndrome virus by DNA Construct Expressing Long Hairpin-RNA Against ICP11 Gene

Design and Construction of Shrimp Antiviral DNA Vaccines Expressing Long and Short Hairpins for Protection by RNA Interference

Contact Info

Product

Resources

About