shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA.

shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Research Abstract Details 

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shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Abstract Text:

Alexander V Vlassov,Brent Korba,Kristine Farrar,Sampa Mukerjee,Attila A Seyhan,Heini Ilves,Roger L Kaspar,Devin Leake,Sergei A Kazakov,Brian H Johnston,

Hepatitis C virus (HCV) is a leading cause of liver cirrhosis and hepatocellular carcinoma worldwide. Currently available treatment options are of limited efficacy, and there is an urgent need for development of alternative therapies. RNA interference (RNAi) is a natural mechanism by which small interfering RNA (siRNA) or short hairpin RNA (shRNA) can mediate degradation of a target RNA molecule in a sequence-specific manner. In this study, we screened in vitro-transcribed 25-bp shRNAs targeting the internal ribosome entry site (IRES) of HCV for the ability to inhibit IRES-driven gene expression in cultured cells. We identified a 44-nt region at the 3'-end of the IRES within which all shRNAs efficiently inhibited expression of an IRES-linked reporter gene. Subsequent scans within this region with 19-bp shRNAs identified even more potent molecules, providing effective inhibition at concentrations of 0.1 nM. Experiments varying features of the shRNA design showed that, for 25-bp shRNAs, neither the size of the loop (4-10 nt) nor the sequence or pairing status of the ends affects activity, whereas in the case of 19-bp shRNAs, larger loops and the presence of a 3'-UU overhang increase efficacy. A comparison of shRNAs and siRNAs targeting the same sequence revealed that shRNAs were of comparable or greater potency than the corresponding siRNAs. Anti-HCV activity was confirmed with HCV subgenomic replicons in a human hepatocyte line. The results indicate that shRNAs, which can be prepared by either transcription or chemical synthesis, may be effective agents for the control of HCV.

shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Publishing Authors By Initials

AV Vlassov,B Korba,K Farrar,S Mukerjee,AA Seyhan,H Ilves,RL Kaspar,D Leake,SA Kazakov,BH Johnston,

For similar investigative techniques: genetic techniques: gene transfer techniques: transfection research abstracts see: investigative techniques: genetic techniques: gene transfer techniques: transfection research

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shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Oligonucleotides

VOLUME: 17

Page Numbers: 223-36

Journal Abbreviation:

ISSN: 1545-4576

DAY: 3

MONTH: 12

YEAR: 2007

shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101188415

shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Keywords Mesh Terms:

KEYWORDS: Transfection

MESH TERMS: genetics

Chemical & Substance for Abstract: shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA. Information

Substance Name: RNA, Viral

Registry Number: 0

Grant and Affiliation Information for shRNAs targeting hepatitis C: effects of sequence and structural features, and comparision with siRNA.

AFFILIATION: SomaGenics, Inc., Santa Cruz, CA 95060, USA.

Country: United States

AGENCY: United States NIAID

GRANT: R44AI056611

ACRONYM: AI

MEDLINETA: Oligonucleotides

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