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
'"/>

Source:


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:
*Name:
*Comment:
*Email:


(Date:6/22/2017)... ... June 21, 2017 , ... RMC Pharmaceutical ... and engages Timothy Reinhardt to manage the new site. , Tim has 25 ... with his most recent role as the Director of Manufacturing and Supplier Quality ...
(Date:6/22/2017)... ... June 22, 2017 , ... For the months of May ... Spotlight series on “Cell Therapy Regulation” for its regenerative medicine followship. ... the unique regulatory challenges of stem cell medical research. , Stem cell clinical ...
(Date:6/22/2017)... Lawrence, MA (PRWEB) , ... June 22, 2017 , ... ... completed ILVO validation. The AMPH test was determined to be appropriate as a screening ... for visual interpretation, on the Charm EZ system, and the Charm EZ Lite system. ...
(Date:6/20/2017)... , June 20, 2017  Kibow Biotech Inc., a ... announce the issuance of a new patent covering a ... by the U.S. Patent and Trademark Office on May ... of the Buzz of Bio award in 2014 in ... to developing non-drug approaches to chronic disease. Renadyl™, the ...
Breaking Biology Technology:
(Date:5/23/2017)...  Hunova, the first robotic gym for the rehabilitation and functional motor ... Genoa, Italy . The first 30 robots will be ... USA . The technology was developed and patented at the ... spin-off Movendo Technology thanks to a 10 million euro investment from entrepreneur ... ...
(Date:5/6/2017)... -- RAM Group , Singaporean based technology ... biometric authentication based on a novel  quantum-state ... perform biometric authentication. These new sensors are based on a ... Group and its partners. This sensor will have widespread ... security. Ram Group is a next generation sensor ...
(Date:4/18/2017)... 2017  Socionext Inc., a global expert in SoC-based imaging and ... the M820, which features the company,s hybrid codec technology. A demonstration ... Probe, Inc., will be showcased during the upcoming Medtec Japan at ... the Las Vegas Convention Center April 24-27. ... Click here for an ...
Breaking Biology News(10 mins):