Introduction
Until recently, gene knockdown or knockout technologies, such as antisense, ribozyme, and gene knockouts were used
to perform loss-of-function studies. However, the post-genomics era calls for high-throughput gene function studies
which the former technologies were unable to answer due to poor reproducibility, high cost, and excessive time to
results. The advent of siRNA technology has opened up many new possibilities in the field of gene suppression.
siRNA Mechanism
siRNA is the term for 20 - 25-base pair RNA duplexes, where the two terminal 3'-nucleotides are unpaired (3' overhang).
When siRNAs are introduced into cells they combine with a protein complex called the RNA-induced silencing complex
(RISC) and are unwound by a helicase. The RISC complex containing single stranded RNA complementary to the target mRNA
then recognizes and binds to the target mRNA. After binding the mRNA, the argonaute protein Ago2 cleaves it and
complete degradation of the target mRNA is carried out by ribonuclease activity (as a result of the lack of protection
by 5' caps or poly (A) tails). This exciting technology is one of the most effective methods for the silencing of
specific target genes and is a must for gene function validation studies, drug target validation, and for gene therapy
studies. siRNA has the following advantages over other RNAi technology:
• Reduced time and costs: Less screening is needed to obtain highly effective siRNA.
• High efficacy at lower concentration: lower concentrations provide effective gene silencing and minimizes off
target effects
• Specificity: siRNA is a highly specific target knockout mechanism based on the natural biological mechanisms of RNAi
Bioneer manufactures high quality and cost-effective siRNA. Every siRNA is produced in an automated high-throughput
RNA production system under clean room conditions and undergo rigorous QC tests.
Features and Benefits
| Performance Guarantee: |
When purchasing 3 siRNAs for the same gene, Bioneer guarantees at least an 80% reduction* in the target mRNA level
for 2 of the three siRNAs.
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| Purification: |
For your more demanding applications, Bioneer's automated HPLC and Bio-RP purification methods ensure high quality,
high- throughput siRNAs at an affordable price.
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| Affordable pricing: |
Bioneer provides a variety of high quality siRNA products at an affordable price.
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| Synthesis and QC: |
Bioneer siRNAs are produced in clean room facility by fully automated high-throughput siRNA production system.
Bioneer siRNA products are assessed by MALDI-TOF Mass spectrometry analysis. Mass spec data is provided with every siRNA.
Additionally, siRNAs are tested by gel electrophoresis to verify that both RNA strands annealed properly.
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All Bioneer siRNAs are provided as double-stranded siRNA. Each sense siRNA and an antisense RNA are QC'ed by MALDI-TOF.
Every annealed siRNA is then QC-tested using PAGE to confirm proper annealing.
Figure 1. MALDI-TOF mass spectrometry analysis of a custom siRNA. All siRNAs are processed by MALDI-TOF
mass spectrometry to ensure its quality.
Figure 2.
PAGE data of double-stranded custom siRNA. Complementary single-stranded RNA strands were hybridized to form siRNA
duplex and analyzed by 15% non-denaturing PAGE.
SS: single-stranded RNA
DS: double-stranded siRNA
Figure 3.
Confocal microscopic image of HeLa cells transfected with FITC-labeled negative control siRNA (Cat No.: SN-1021).
The fluorescent cells indicate that the HeLa cells were successfully transfected with the siRNA.
Figure 4.
Effects of Human GAPDH Positive Control siRNA. HeLa cells were transfected separately with AccuTarget Human GAPDH
Positive Control and Negative Control siRNA using Lipofectamine 2000 (Invitrogen) at a final concentration of 100 nM.
Total cellular RNA was isolated from cells 24 hours after transfection and subjected to Northern blot and Real-Time
PCR analysis. As can be seen from Fig. 1-B, about 3% GAPDH mRNA remained.
Contact
Bioneer, Inc.
Phone: 1-877-264-4300
To place an order: order.usa@bioneer.us.com
Email: support@bioneer.us.com
Notice to Purchaser
All siRNA Products: For Research Use Only. Not For Use in Diagnostic Procedures.
Limited License
This product is licensed under European Patents 1144623, 121945 and foreign equivalents from Alnylam Pharmaceuticals,
Inc., Cambridge, USA and is provided only for use in academic and commercial research whose purpose is to elucidate gene
function, including research to validate potential gene products and pathways for drug discovery and development and to
screen non-siRNA based compounds (but excluding the evaluation or characterization of this product as the potential
basis for a siRNA-based drug) and not for any other commercial purposes. Information about licenses for commercial
use (including discovery and development of siRNA-based drugs) is available from Alnylam Pharmaceuticals, Inc.,
300 Third Street, Cambridge, MA 02142, USA.
This product is sold for research use only and is not to be administered to humans or used for medical diagnostics.
Buyer acknowledges and agrees that all intellectual property rights in the products (including, without limitation,
the siRNA sequences used to create such products) and in any Bioneer technology, intellectual property and know-how
used to make or useful for the manufacture or use of the products will at all times remain vested in Bioneer
(other than any ownership interest that buyer may have in non-public proprietary target genes supplied by buyer to
Bioneer in connection with custom products).
Trademark: AccuTarget is a trademark of Bioneer Corporation.