About: Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70

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About: Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70 Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70
Creator	Liu, Mingqiu Yan, Weiyao Zheng, Zhaoxin Liu, Shuang Qi, Zhongtian Xiao, An Bian, Zhongqi Cao, Mingmei Jiao, Ye
Source	Medline; PMC
abstract	BACKGROUND: Hepatitis B virus (HBV) infection is a major health concern with more than two billion individuals currently infected worldwide. Because of the limited effectiveness of existing vaccines and drugs, development of novel antiviral strategies is urgently needed. Heat stress cognate 70 (Hsc70) is an ATP-binding protein of the heat stress protein 70 family. Hsc70 has been found to be required for HBV DNA replication. Here we report, for the first time, that combined siRNAs targeting viral gene and siHsc70 are highly effective in suppressing ongoing HBV expression and replication. METHODS: We constructed two plasmids (S1 and S2) expressing short hairpin RNAs (shRNAs) targeting surface open reading frame of HBV(HBVS) and one plasmid expressing shRNA targeting Hsc70 (siHsc70), and we used the EGFP-specific siRNA plasmid (siEGFP) as we had previously described. First, we evaluated the gene-silencing efficacy of both shRNAs using an enhanced green fluorescent protein (EGFP) reporter system and flow cytometry in HEK293 and T98G cells. Then, the antiviral potencies of HBV-specific siRNA (siHBV) in combination with siHsc70 in HepG2.2.15 cells were investigated. Moreover, type I IFN and TNF-α induction were measured by quantitative real-time PCR and ELISA. RESULTS: Cotransfection of either S1 or S2 with an EGFP plasmid produced an 80%–90% reduction in EGFP signal relative to the control. This combinational RNAi effectively and specifically inhibited HBV protein, mRNA and HBV DNA, resulting in up to a 3.36 log(10) reduction in HBV load in the HepG2.2.15 cell culture supernatants. The combined siRNAs were more potent than siHBV or siHsc70 used separately, and this approach can enhance potency in suppressing ongoing viral gene expression and replication in HepG2.2.15 cells while forestalling escape by mutant HBV. The antiviral synergy of siHBV used in combination with siHsc70 produced no cytotoxicity and induced no production of IFN-α, IFN-β and TNF-α in transfected cells. CONCLUSIONS: Our combinational RNAi was sequence-specific, effective against wild-type and mutant drug-resistant HBV strains, without triggering interferon response or producing any side effects. These findings indicate that combinational RNAi has tremendous promise for developing innovative therapy against viral infection.
has issue date	2012-11-16(xsd:dateTime)
bibo:doi	10.1186/1743-422x-9-275
bibo:pmid	23158906
has license	cc-by
sha1sum (hex)	a94e4109b44346c490243c71fd533cd1a9fd047c
schema:url	https://doi.org/10.1186/1743-422x-9-275
resource representing a document's title	Anti-HBV efficacy of combined siRNAs targeting viral gene and heat shock cognate 70
has PubMed Central identifier	PMC3534549
has PubMed identifier	23158906
schema:publication	Virol J
resource representing a document's body	covid:a94e4109b44346c490243c71fd533cd1a9fd047c#body_text
is schema:about of	named entity 'Heat stress' named entity 'health concern' named entity 'plasmids' named entity 'effective' named entity 'HBV' covid:arg/a94e4109b44346c490243c71fd533cd1a9fd047c named entity 'HBV' named entity 'plasmid' named entity 'RT-PCR' named entity 'Invitrogen' named entity 'HBV' named entity 'siRNA' named entity 'gene silencing' named entity 'RNAi' named entity 'RT-PCR' named entity 'siRNAs' named entity 'HBV' named entity 'mRNAs' named entity 'siRNA' named entity 'protein' named entity 'EGFP' named entity 'viral persistence' named entity 'viral proteins' named entity 'virus' named entity 'virus' named entity 'siRNA' named entity 'HBV' named entity 'EGFP' named entity 'polo-like kinase 1' named entity 'intracellular' named entity 'plasmids' named entity 'transfected' named entity 'subcloned' named entity '3.5' named entity 'EGFP' named entity 'gene product' named entity 'EGFP' named entity 'EGFP' named entity 'protein synthesis' named entity 'HBV' named entity 'virus' named entity 'transfected' named entity 'transfection' named entity 'HBV' named entity 'RNAs' named entity 'BamHI' named entity 'flow cytometry' named entity 'HBV' named entity 'miR-122' named entity 'Millipore' named entity 'National Center for Biotechnology Information' named entity 'France' named entity 'IFN' named entity 'open reading frames' named entity '3.5' named entity 'IFN-β' named entity 'shRNA' named entity 'mammalian cells' named entity 'protein' named entity 'development' named entity 'potencies' named entity 'shRNA' named entity 'mRNA' named entity 'shRNAs' named entity 'genotypes' named entity 'TNF-α' named entity 'monoclonal antibodies' named entity 'mutation' named entity 'HBV' named entity 'virus' named entity 'Pol II' named entity 'protein expression' named entity 'sequence data'

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