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 25, 2015 , ... Jessica Richman and Zachary ... in their initial angel funding process. Now, they are paying it forward to ... early stage investments in the microbiome space. In this, they join other ...
(Date:11/24/2015)... VANCOUVER , Nov. 24, 2015 /CNW/ - iCo ... ICOTF), today reported financial results for the quarter ... are expressed in Canadian dollars and presented under ... the United States ," said Andrew ... "These advancements regarding iCo-008 are not only value ...
(Date:11/24/2015)... 24, 2015 --> ... report "Oligonucleotide Synthesis Market by Product & Services (Primer, ... Diagnostic, DNA, RNAi), End-User (Research, Pharmaceutical & Biotech, Diagnostic ... the market is expected to reach USD 1,918.6 Million ... a CAGR of 10.1% during the forecast period. ...
(Date:11/24/2015)...  Tikcro Technologies Ltd. (OTCQB: TIKRF) today announced that its Annual General ... a.m. Israel time, at the law offices of ... 36 th Floor, Tel Aviv, Israel . ... Izhak Tamir to the Board of Directors; , election ... , approval of an amendment to certain terms of options granted to ...
Breaking Biology Technology:
(Date:10/29/2015)... Calif. , Oct. 29, 2015  The J. ... new report titled, "DNA Synthesis and Biosecurity: Lessons Learned ... the Department of Health and Human Services guidance for ... in 2010. --> ... it also has the potential to pose unique biosecurity ...
(Date:10/29/2015)... ANN ARBOR, Mich. , Oct. 29, 2015 ... with Eurofins Genomics for U.S. distribution of its ... DNA-seq kit and Rubicon,s new ThruPLEX Plasma-seq ... DNA to enable the preparation of NGS libraries ... in plasma for diagnostic and prognostic applications in ...
(Date:10/29/2015)... 29, 2015 NXTD ) ... focused on the growing mobile commerce market and ... StackCommerce, a leading marketplace to discover and buy ... smart wallet on StackSocial for this holiday season. ... the "Company"), a biometric authentication company focused on ...
Breaking Biology News(10 mins):