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Inhibition of core gene of HCV 3a genotype using synthetic and vector derived siRNAs.
Virol J. 2010 Nov 13; 7:318.VJ

Abstract

BACKGROUND

Hepatitis C virus (HCV) is a major causative agent of liver associated diseases throughout the world, with genotype 3a responsible for most of the cases in Pakistan. Due to the limited efficiency of current therapy, RNA interference (RNAi) a novel regulatory and powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process represents an alternative option.

RESULTS

The current study was purposed to assess and explore the possibility of RNAi to silence the HCV-3a Core gene expression, which play complex role in regulation of cell growth and host genes expression essential for infectivity and disease progression. To identify the potent siRNA target sites, 5 small interfering RNAs (siRNAs) against Core gene were designed and in vitro transcribed after consensus sequence analysis of different HCV-3a isolates. Antiviral effects of siRNAs showed upto 80% inhibition of Core gene expression by different siRNAs into Huh-7 cells as compared with Mock transfected and control siRNAs treated cells. For long lasting effect of siRNAs, vector based short hairpin siRNAs (shRNAs) were designed and tested against HCV-3a Core which resulted in a similar pattern of inhibition on RNA and protein expression of HCV Core as synthetic siRNAs. Furthermore, the efficacy of cell culture tested siRNA and shRNA, were evaluated for inhibition of HCV replication in HCV infected serum inoculated Huh-7 cells and a significant decrease in HCV viral copy number was observed.

CONCLUSIONS

Our results support the possibility of using consensus siRNA and shRNA-based molecular therapy as a promising strategy in effective inhibition of HCV-3a genotype.

Authors+Show Affiliations

Applied and Functional Genomics Laboratory, National Center of Excellence in Molecular Biology, University of Punjab, Lahore 53700, Pakistan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21073745

Citation

Khaliq, Saba, et al. "Inhibition of Core Gene of HCV 3a Genotype Using Synthetic and Vector Derived SiRNAs." Virology Journal, vol. 7, 2010, p. 318.
Khaliq S, Jahan S, Ijaz B, et al. Inhibition of core gene of HCV 3a genotype using synthetic and vector derived siRNAs. Virol J. 2010;7:318.
Khaliq, S., Jahan, S., Ijaz, B., Ahmad, W., Asad, S., Pervaiz, A., Samreen, B., Khan, M., & Hassan, S. (2010). Inhibition of core gene of HCV 3a genotype using synthetic and vector derived siRNAs. Virology Journal, 7, 318. https://doi.org/10.1186/1743-422X-7-318
Khaliq S, et al. Inhibition of Core Gene of HCV 3a Genotype Using Synthetic and Vector Derived SiRNAs. Virol J. 2010 Nov 13;7:318. PubMed PMID: 21073745.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Inhibition of core gene of HCV 3a genotype using synthetic and vector derived siRNAs. AU - Khaliq,Saba, AU - Jahan,Shah, AU - Ijaz,Bushra, AU - Ahmad,Waqar, AU - Asad,Sultan, AU - Pervaiz,Asim, AU - Samreen,Baila, AU - Khan,Mahwish, AU - Hassan,Sajida, Y1 - 2010/11/13/ PY - 2010/08/10/received PY - 2010/11/13/accepted PY - 2010/11/16/entrez PY - 2010/11/16/pubmed PY - 2011/2/1/medline SP - 318 EP - 318 JF - Virology journal JO - Virol. J. VL - 7 N2 - BACKGROUND: Hepatitis C virus (HCV) is a major causative agent of liver associated diseases throughout the world, with genotype 3a responsible for most of the cases in Pakistan. Due to the limited efficiency of current therapy, RNA interference (RNAi) a novel regulatory and powerful silencing approach for molecular therapeutics through a sequence-specific RNA degradation process represents an alternative option. RESULTS: The current study was purposed to assess and explore the possibility of RNAi to silence the HCV-3a Core gene expression, which play complex role in regulation of cell growth and host genes expression essential for infectivity and disease progression. To identify the potent siRNA target sites, 5 small interfering RNAs (siRNAs) against Core gene were designed and in vitro transcribed after consensus sequence analysis of different HCV-3a isolates. Antiviral effects of siRNAs showed upto 80% inhibition of Core gene expression by different siRNAs into Huh-7 cells as compared with Mock transfected and control siRNAs treated cells. For long lasting effect of siRNAs, vector based short hairpin siRNAs (shRNAs) were designed and tested against HCV-3a Core which resulted in a similar pattern of inhibition on RNA and protein expression of HCV Core as synthetic siRNAs. Furthermore, the efficacy of cell culture tested siRNA and shRNA, were evaluated for inhibition of HCV replication in HCV infected serum inoculated Huh-7 cells and a significant decrease in HCV viral copy number was observed. CONCLUSIONS: Our results support the possibility of using consensus siRNA and shRNA-based molecular therapy as a promising strategy in effective inhibition of HCV-3a genotype. SN - 1743-422X UR - https://www.unboundmedicine.com/medline/citation/21073745/Inhibition_of_core_gene_of_HCV_3a_genotype_using_synthetic_and_vector_derived_siRNAs_ L2 - https://virologyj.biomedcentral.com/articles/10.1186/1743-422X-7-318 DB - PRIME DP - Unbound Medicine ER -