Call +49 (0)7250 33 13 400, email value@bts-biotech.com

EXCELLENT TOOLS & SERVICES FOR DNA, RNA, PROTEIN OR CELL RESEARCH

AccuTarget™ Fluorescein-labeled Control miRNAs


  • For RNAi research BIONEER offers:

  • Predesigned siRNA libraries

  • PCR primers for knockdown validation

  • Human validated siRNA libraries

  • Human siRNA library sets and subsets

  • Control siRNAs

  • siRNA design and synthesis 

  • Below given product list is an example for general price overview

  • To order your RNAi, please refer to BIONEER online ordering or e-mail bts

Inquire prices at value@bts-biotech.com

bioneer

Request a Sample.

Description

BIONEER is currently the market leader in RNAi technology in Asia. Company's RNAi portfolio consists of six areas:

  • Predesigned siRNA libraries
  • PCR primers for knockdown validation
  • Human validated siRNA libraries
  • Human siRNA library sets and subsets
  • Control siRNAs
  • siRNA design and synthesis


All Bioneer's siRNAs are designed using proprietary Turbo si-Designer software. This software identifies highly effective siRNA target sites with remarkably high knockdown rates. Critical parameters including base composition, thermodynamic instability and base preference are all considered in the design algorithm. siRNAs spanning SNP sites are removed and finally, non-specific siRNAs are eliminated following homology searching by BLAST and Smith-Waterman algorithms. The result is an siRNA with extraordinary knockdown efficiency and minimal off-target effects.We are so confident in our software that we have the best guarantee in the industry – Buy three of our AccuTarget™ siRNAs, and if 2 of them don’t have knockdown rates of at least 80%, we will replace them free of charge!

The Bioneer 80% 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 3 siRNAs. If there is not a > 80% reduction in the mRNA level of the target gene, Bioneer will supply 2 siRNAs free of charge.*

*Bioneer reserves the right to request supporting data inclusive of:
1. Transfection efficiency data: NC (AccuTarget™ Fluorescein-labeled Negative Control) and siRNA concentration at 100 nM)
2. siRNA knockdown efficiency data: PC (AccuTarget™ GAPDH/GFP/Luciferase siRNA) and NC (AccuTarget™ Negative Control)

AccuTarget™ miRNA mimic controls

We offer AccuTarget™ miRNA mimic controls to optimize assay conditions for miRNA mimic function studies. Both positive and negative controls are provided for miRNA gain-of-function studies using Bioneer's AccuTarget™ miRNA mimics.

AccuTarget™ miRNA housekeeping Positive controls target the 3' UTR (untranslated region) of the standard housekeeping gene, GAPDH, and BIONEER's miRNA mimic Negative controls' sequences are based on common miRNA structure for use as negative experimental controls in human, mouse, and rat cells. The negative controls have been analyzed by BLAST against all human, mouse and rat genomic sequences and miRNA sequences in the current miRBase Database. BIONEER offers two universal negative controls for mimics. In addition, AccuTarget™ miRNA control sets consisting of a Positive and two Negative miRNA controls are also available for user convenience.

 MicroRNAs (miRNAs) are 21-25 nucleotide(nt)-long single-stranded RNA molecules that serve as a post-transcriptional regulator of gene expression in eukaryotes. The human genome may encode over 1000 miRNAs, which bind with imperfect complementarity to their target mRNAs, generally within the 3’UTR (untranslated region), and repress protein production by destabilizing the mRNA as well as translational suppression. miRNA-mediated translational repression has important roles in wide range of biological process, including development, cell proliferation and differentiation, apoptosis and metabolism [1].

MicroRNA Pathway

The biogenesis of miRNAs consists of two sequential processing events. Primary miRNA transcripts (pri-miRNAs), which contain one or multiple stem-loop hairpin structures, are mostly derived from Pol II-mediated transcription. In first step towards the canonical miRNA maturation pathway, pri-miRNA is cleaved by the microprocessor complex, RNase Ⅲ enzyme Drosha, to yield the pre-miRNA, a hairpin-shaped intermediate precursor ~70 nt in length. Pre-miRNAs are then exported from the nucleus to the cytoplasm by Exprotin5, where another RNaseⅢ enzyme Dicer catalyzes the second processing event for miRNA biogenensis and liberates the mature miRNA duplexes. The mature miRNA duplexes consist of the mature miRNA strand and the miRNA* strand, which are derived from two separate arms of the hairpin stem within the miRNA precursor. The miRNA is loaded into an Argonaute-containing RNA-induced silencing complex (RISC), whereas the miRNA* strand is typically degraded. The Ago:miRNA complex then dissociates from RISC loading complex, and become the core of the RISC complex to regulate post-transcriptional gene repression of specific target mRNAs[2].



A recently identified extensive class of small RNAs, called miRNA, has provided new insights in biotechnology. Although they were discovered and recognized relatively recently, miRNAs have been recognized as the most important gene regulators at the post-transcriptional level, and several studies indicated that miRNAs regulate the expression of more than 30 % protein coding genes [3]. The accumulating knowledge about their biogenesis, gene expression regulation mechanism and functions will add a new dimension to our understanding about the complex gene regulatory networks. Recent investigations demonstrate that miRNAs have a unique expression profiles in different cancer types at different stages and play an important role in many disease and viral infections. These results suggest that miRNAs can function as a novel biomarker for disease diagnosis and perform a new strategy for miRNA gene therapy [4].

 

1. Sci china Ser C-Life Sci, 2009, 52(4):323-330
2. Annu. Rev. Cel. Dev. Biol. 2007. 23:175-205
3. Virchows Arch. 2008. 452:1-10
4. J. Cell. Mol. Med. 2008. 12(1): 3-21

Performance of miRNA mimic Positive and Negative controls



Figure1 . AccuTarget™ miRNA Positive & Negative controls were transfected at 20 nM using Lipofectamine™ RNAiMAX into HeLa cell lines and assessed for their ability to decrease target mRNA levels. Down-regulation of GAPDH was determined using the real-time quantitative RT-PCR at 48 hours post-transfection using Bioneer's Exicylcer™ 96 qPCR instrument.

Specifications

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



sirna mechanism

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.

 

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.

rnai figure2

Figure 1. MALDI-TOF mass spectrometry analysis of a custom siRNA. All siRNAs are processed by MALDI-TOF mass spectrometry to ensure its quality.


rnai figure3

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


rnai figure4

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.


rnai figure5

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: +82-42-930-8777
To place an order: sales@bioneer.com
Email: sales@bioneer.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.

Supporting Data

References

Resources

Manual

• miRNA mimic and inhibitor user manual

• Protocol for siRNA annealing

• siRNA User Manual

• qPCR User Manual

 

Brochure

• siRNA Products and services 2010

 

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

You may also be interested in the following product(s)