Navigation Links
Your Data: Silencing Hepatitis C Virus Replication With In Vitro Transcribed siRNAs

The Silencer siRNA Construction Kit uses an in vitro transcription reaction and column purification to prepare transfection-ready siRNAs. With this kit, multiple siRNAs can be prepared in ~24 hours. Below is one example of how siRNAs prepared by the Silencer siRNA Construction Kit were used in gene silencing studies.

Kapadia S, Brideau-Anderson A, and Chisari FV (2003) Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc Natl Acad Sci USA 100: 2014-2018.

Dr. Sharookh B. Kapadia and colleagues at the Scripps Research Institute used siRNAs prepared with the Silencer siRNA Construction Kit to demonstrate that RNAi can be used to inhibit Hepatitis C Virus (HCV) replication. The researchers generated multiple siRNAs targeting various regions of the HCV subgenomic replicon sequence. These siRNAs were transfected into an Huh-7 cell line that stably expressed the HCV RNA replicon and screened for their gene silencing activities by real-time RT-PCR. Two of the siRNAs, NS3-1948 and NS5B-6133, showed the greatest specific inhibition of HCV RNA replication and were tested further for their ability to suppress HCV RNA and protein expression. As shown in Figure 1, HCV protein expression levels were significantly reduced on Day 4 (Figure 1B) and Day 6 (Figure 1C) post-transfection. Similar results are seen at the mRNA level (Figure 2). Viral replication was also inhibited. These results suggest that RNAi might represent a new approach for the treatment of HCV infection.

Figure 1. The Effect of RNAi on HCV Protein Expression in S11791 Cells. Western blot analysis was performed on total cell lysates harvested from mock- or siRNA-transfected S11791 cells at various days post-transfection by using NS3- and NS5B-specific antibodies. Molecular mass markers (in kilodaltons) are shown to the right of each gel. Shown are representative data of three independent experiments.

Figure 2. Effect of HCV-specific siRNAs on HCV and GAPDH RNA Transcript Levels. Total RNA was isolated from S11791 cells that were transfected with siRNA specific for NS3, NS5B, GAPDH, or an irrelevant scrambled (scr) siRNA 2, 4, and 6 days previously. Equal amounts of RNA were separated on a denaturing agarose gel, transferred to membrane and hybridized with probes to HCV and GAPDH. GAPDH signal was used for normalization of the HCV signal.

back to top

Ordering Information
Cat# Product Name Size 1620 Silencer siRNA Construction Kit 15 siRNA synthesis rxns


Page: All 1 2

Related biology technology :

1. Your Data: Detection of Oocyte-specific Gene Expression Using Ambions Cells-to-cDNA Kit
2. Your Data: Use of pSilencer 1.0-U6 siRNA Expression Vector in the Analysis of Synaptotagmin Location and Function
3. Your Data: RNAlater Stories
4. Your Data: Silencing Neural Cell Adhesion Genes with In Vitro Transcribed siRNAs
5. Your Data: Silencing of a Pro-apoptotic Transcription Factor Using Synthetic siRNA
6. RNAi Gene Silencing System
7. Quantification of siRNA Silencing Efficiency Using the LightCycler System
8. Measuring Gene Silencing Effects by RT-PCR Without RNA Isolation
9. Drive Long-Term Silencing with CMV Vectors
10. Enhanced siRNA Delivery and Long-term Gene Silencing
11. Fast, Accurate Assessment of siRNA-induced Gene Silencing
Post Your Comments:

(Date:11/25/2015)... , November 26, 2015 ... Biobanking Market 2016 - 2020 report analyzes that ... integrity and quality in long-term samples, minimizing manual ... cost-effectiveness. Automation minimizes manual errors such as mislabeling ... efficiency. Further, it plays a vital role in ...
(Date:11/25/2015)... , Nov. 25, 2015  Neurocrine Biosciences, Inc. ... Gorman , President and CEO of Neurocrine Biosciences, will ... Conference in New York . ... visit the website approximately 5 minutes prior to the ... replay of the presentation will be available on the ...
(Date:11/25/2015)... ... November 25, 2015 , ... Jessica ... on AngelList early in their initial angel funding process. Now, they are paying ... looking to make early stage investments in the microbiome space. In this, ...
(Date:11/24/2015)... /CNW/ - iCo Therapeutics ("iCo" or "the Company") (TSX-V: ... the quarter ended September 30, 2015. Amounts, unless ... presented under International Financial Reporting Standards ("IFRS"). ... Andrew Rae , President & CEO of ... only value enriching for this clinical program, but ...
Breaking Biology Technology:
(Date:11/20/2015)... OXFORD, Connecticut , November 20, 2015 ... biometric authentication company focused on the growing mobile commerce ... its CEO, Gino Pereira , was recently interviewed ... The interview will air on this weekend on ... Bloomberg Latin America . --> NXTD ) ...
(Date:11/17/2015)... 17, 2015 Paris from ... --> Paris from 17 th until ... biometrics innovation leader, has invented the first combined scanner in ... same scanning surface. Until now two different scanners were required: ... can capture both on the same surface. This innovation ...
(Date:11/12/2015)...  A golden retriever that stayed healthy despite having ... provided a new lead for treating this muscle-wasting disorder, ... of MIT and Harvard and the University of São ... Cell, pinpoints a protective gene that boosts ... The Boston Children,s lab of Lou Kunkel , ...
Breaking Biology News(10 mins):