Inhibition of Respiratory Syncytial Virus Replication by Simultaneous Targeting of mRNA and Genomic RNA Using Dual-Targeting siRNAs

Malekshahi, Somayeh Shatizadeh; Salimi, Vahid; Arefian, Ehsan; Fatemi-Nasab, Ghazal; Adjaminejad-Fard, Sarvin; Yavarian, Jila; Mokhtari‐Azad, Talat

doi:10.1007/s12033-016-9976-4

Cited by 7 publications

(6 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As shown in the present report, silencing HRSV N drastically reduced the innate immune response, as well as the expression of F and G. On the contrary, silencing F had a moderate effect on the innate immune response but drastically reduced the expression of the protein F, which is the main target for neutralizing antibodies. Previous studies using siRNAs against HRSV have focused on proteins involved in replication/transcription (N, P, and M2-2) [26,27,30,32,34] or the fusion F protein [30,33]. In all cases, a substantial decrease in virus titers has been reported.…”

Section: Discussionmentioning

confidence: 99%

“…Pioneering studies showing the efficacy of siRNAs against viruses included HRSV [21], human immunodeficiency virus-1 (HIV-1) [22], and the influenza virus [23,24]. Afterward, several studies have reported the inhibition of HRSV by siRNA-mediated silencing of a number of HRSV genes, including NS1 , N , P , F, and M2-2 [25,26,27,28,29,30,31,32,33,34]. Some of them have been tested in vivo in the mouse model and found to restrict HRSV replication [25,27,32].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication

2019

View full text Add to dashboard Cite

Human respiratory syncytial virus (HRSV) infection is a common cause of severe lower respiratory tract diseases such as bronchiolitis and pneumonia. Both virus replication and the associated inflammatory immune response are believed to be behind these pathologies. So far, no vaccine or effective treatment is available for this viral infection. With the aim of finding new strategies to counteract HRSV replication and modulate the immune response, specific small interfering RNAs (siRNAs) were generated targeting the mRNA coding for the viral fusion (F) protein or nucleoprotein (N), or for two proteins involved in intracellular immune signaling, which are named tripartite motif-containing protein 25 (TRIM25) and retinoic acid-inducible gene-I (RIG-I). Furthermore, two additional bispecific siRNAs were designed that silenced F and TRIM25 (TRIM25/HRSV-F) or N and RIG-I (RIG-I/HRSV-N) simultaneously. All siRNAs targeting N or F, but not those silencing TRIM25 or RIG-I alone, significantly reduced viral titers. However, while siRNAs targeting F inhibited only the expression of the F mRNA and protein, the siRNAs targeting N led to a general inhibition of viral mRNA and protein expression. The N-targeting siRNAs also induced a drastic decrease in the expression of genes of the innate immune response. These results show that both virus replication and the early innate immune response can be regulated by targeting distinct viral products with siRNAs, which may be related to the different role of each protein in the life cycle of the virus.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication

2019

View full text Add to dashboard Cite

show abstract

“…New strategies such as second-generation siRNAs against the paramyxoviral RNA polymerase large subunit and a siRNA cocktail against influenza virus have also been applied [51]. In a recent study using dual-targeting siRNAs, which can knock down the expression of mRNA and viral genomic RNA simultaneously with its two active strands, the replication of respiratory syncytial virus was more effectively inhibited [52].…”

Section: Therapeutic Applications Of Rnaimentioning

confidence: 99%

RNA interference-based therapy and its delivery systems

Chen

Mangala

Rodríguez-Aguayo

et al. 2017

Cancer Metastasis Rev

233

164

View full text Add to dashboard Cite

RNA interference (RNAi) is considered a highly specific approach for gene silencing and holds tremendous potential for treatment of various pathologic conditions such as cardiovascular diseases, viral infections, and cancer. Although gene silencing approaches such as RNAi are widely used in preclinical models, the clinical application of RNAi is challenging primarily because of the difficulty in achieving successful systemic delivery. Effective delivery systems are essential to enable the full therapeutic potential of RNAi. An ideal nanocarrier not only addresses the challenges of delivering naked siRNA/miRNA, including its chemically unstable features, extracellular and intracellular barriers, and innate immune stimulation, but also offers "smart" targeted delivery. Over the past decade, great efforts have been undertaken to develop RNAi delivery systems that overcome these obstacles. This review presents an update on current progress in the therapeutic application of RNAi with a focus on cancer therapy and strategies for optimizing delivery systems, such as lipid-based nanoparticles.

show abstract

“…After verifying the accuracy of cloning by sequencing, the accurate clones were used for transfection. As a mock (no shRNA GFP lentivector),we used pEZX-MR03 that contained the EGFP gene under the control of a CMV promoter lacking any significant shRNA sequences [33].…”

Section: Production Of Lentivector Expressing Shrnas and Orfv-dna Polmentioning

confidence: 99%

Lentivirus expressing shRNAs inhibit the replication of contagious ecthyma virus by targeting DNA polymerase gene

et al. 2020

Self Cite

View full text Add to dashboard Cite

Background: Contagious ecthyma or Orf is known as a zoonotic disease remains prevalently worldwide despite the application of some control strategies against it. RNAi particularly shRNA provides us with the chance to tackle this obstacle by an encouraging new approach. The current study indicates the design and experiment of thirdgeneration lentivirus packaging systems delivering shRNAs to inhibit Orf virus (ORFV) replication and infection. Given the importance of DNA-pol gene in virus replication, in this study, three shRNAs against this gene were designed and cloned into lentiviral vectors to stabilize the expression of shRNAs. After producing lentivectors expressing ORFV-DNA-pol in HEK293T cells, the synthesized shRNAs were applied to downregulate viral replication and gene expression. The reduction in viral titer and RNA was evaluated by TCID50 test as well as real-time RT-PCR. The results were then analyzed in comparison with the control group. Results: Designed shRNAs significantly reduced virus yield approximately 90 to 97% and 96.8 to 99.4%, respectively compared to the control groups (cells infected with ORFV and infected with ORFV and scrambled vector) by TCID50 test. Real-time RT-PCR revealed a dramatic reduction in the expression of viral RNA approximately 99% compared to cells infected with ORFV and from 92.6 to 99%, respectively compared to cells infected with ORFV and scrambled vector. Conclusions: Therefore, it can be stated that RNAi is capable of being used as a potent therapeutically option against viruses like ORFV.

show abstract

Inhibition of Respiratory Syncytial Virus Replication by Simultaneous Targeting of mRNA and Genomic RNA Using Dual-Targeting siRNAs

Cited by 7 publications

References 33 publications

siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication

siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication

RNA interference-based therapy and its delivery systems

Lentivirus expressing shRNAs inhibit the replication of contagious ecthyma virus by targeting DNA polymerase gene

Contact Info

Product

Resources

About