RNAi
RNAi
This document provides a comprehensive overview of Bioneer's siRNA and miRNA synthesis services, focusing on the AccuTarget™ siRNA products and the Turbo si-Designer algorithm.
AccuTarget™ siRNAs are designed for gene knockdown studies, offering advantages such as reduced time and costs, high efficacy at lower concentrations, and specificity. The siRNA mechanism involves RNA duplexes that bind to the RNA-induced silencing complex (RISC) to degrade target mRNA, making it effective for gene silencing and validation studies.
Bioneer's proprietary Turbo si-Designer algorithm, developed with the National Genome Information Center, identifies effective siRNA target sites. It considers parameters like base composition and thermodynamic instability to minimize off-target effects. The algorithm's efficacy is validated through Real-Time PCR analysis, showing high knockdown rates.
This library offers over 132,000 predesigned siRNAs for more than 44,000 target genes. Bioneer guarantees at least an 80% reduction in target mRNA levels for two out of three siRNAs purchased for the same gene. The library is suitable for functional genomics, gene expression studies, and array analysis.
The primer library consists of 11,154 validated primer sets for human genes, designed for specific and sensitive Real-Time PCR results. It is categorized by gene function and pathway, offering competitive pricing.
Control siRNAs include positive controls for high knockdown rates and negative controls that do not target known genes. These controls are available in fluorescently labeled sets for monitoring transfection efficiency.
The document provides detailed ordering information for various siRNA and primer products, including purification methods and guaranteed yields.
The document lists various AccuTarget™ Negative Control siRNA products, available in different nmole quantities and purification methods (HPLC and Bio-RP). These are used as controls in experiments to ensure the accuracy of siRNA-mediated gene silencing.
Bioneer offers a wide range of premade siRNA sets, including 25,368 predesigned siRNAs and 25 pathway-specific sets. These are available in various nmole yields and are designed for immediate use in experiments related to cellular processes, cancer, and disease research. The document highlights the benefits of these sets, such as pre-validation and competitive pricing.
The document describes AccuTarget™ Real-Time PCR Primer Sets, which are optimized for use with premade siRNAs in real-time PCR applications. These primers are categorized by pathway or gene function and are designed to provide specific and sensitive results.
Bioneer offers custom siRNA synthesis services, allowing researchers to design siRNAs according to their specific needs. The service includes various modification options and guarantees high-quality siRNA production, with quality control measures such as MALDI-TOF mass spectrometry and PAGE analysis.
The document includes a comprehensive FAQ section addressing common questions about siRNA orders, storage, and experimental conditions. It also provides detailed protocols for siRNA dilution and transfection, ensuring proper handling and use of the products.
Figures in the document demonstrate the knockdown efficiency of AccuTarget™ siRNA libraries, with significant percentages of siRNAs achieving high knockdown rates in HeLa cells. This data underscores the effectiveness of Bioneer's siRNA design and validation processes.
1. Plate 3.0×105 HeLa cells per well in a 6-well plate with 2.5 ml growth medium without antibiotics, ensuring 50-60% confluence at transfection time.
2. Before transfection, replace the growth medium with 500 μl fresh medium without serum.
3. Prepare siRNA duplex-Lipofectamine™ RNAiMAX complexes:
a. Dilute siRNA duplex to a final concentration of 5 nM~100 nM in 250 μl medium without serum.
b. Dilute 3.5 μl Lipofectamine™ RNAiMAX in 250 μl medium without serum, incubate for 5 minutes.
c. Combine diluted siRNA duplex with Lipofectamine™ RNAiMAX, mix, and incubate for 20 minutes.
4. Add the mixture to each well, resulting in a total volume of 1 ml, and mix gently.
5. Incubate cells for 5-6 hours at 37˚C in a CO2 incubator.
6. Replace the medium with fresh serum-containing medium and incubate for 24-48 hours before gene knockdown assay.
Table 2 provides the relative surface area and volume of plating medium for various culture vessels, ranging from 96-well plates to 100 mm dishes.
MicroRNAs (miRNAs) are 21-25 nucleotide long RNA molecules that regulate gene expression post-transcriptionally. They play roles in development, cell proliferation, differentiation, apoptosis, and metabolism. The biogenesis involves two processing events, resulting in mature miRNA duplexes that regulate gene repression.
Bioneer offers chemically synthesized miRNA mimics and inhibitors for human miRNAs. These products are available in various scales and are designed for transfection to study gene function. They are purified using HPLC and Bio-RP methods, ensuring high quality and affordability.
AccuTarget™ provides miRNA mimic controls for optimizing assay conditions. Positive controls target GAPDH, while negative controls are designed for minimal sequence identity with human, mouse, and rat miRNAs. These controls are available with fluorescence labeling for monitoring transfection efficiency.
SAMiRNA™ is a novel siRNA delivery system developed by Bioneer, designed for in vivo applications. It involves a single-molecular synthetic siRNA conjugated with lipid and polymer, forming stable nanoparticles for systemic delivery. This technology aims to overcome delivery challenges in RNAi-based therapeutics.
SAMiRNA™ is a novel RNAi prodrug technology designed for therapeutic applications, particularly in cancer and lung fibrosis models. It is characterized by its negligible toxicity, high serum stability, and effective gene silencing capabilities without inducing an innate immune response. This is attributed to Bioneer's expertise in nucleic acid chemistry and manufacturing.
SAMiRNA™ products are intended for research purposes only and are not suitable for diagnostic or therapeutic use in humans. They are protected under U.S. patents and require a license for commercial use.
- Custom siRNA Synthesis: Offers up to 30-mer siRNA with various modifications for specificity, provided purified and ready for animal testing.
- Genome-wide siRNA Library: Available for mouse, rat, and human, designed using the Turbo-si algorithm for high knockdown efficiency.
- Nanoparticle Synthesis: SAMiRNA™ nanoparticles are designed for in vivo delivery, overcoming traditional siRNA delivery challenges.
- In vitro Screening: Validates siRNA efficiency through RT-qPCR, optimizing lead candidates before shipment.
- qPCR Array Service: Quantifies gene knockdown using real-time RT-qPCR, adhering to MIQE guidelines.
Custom services are available through a dedicated ordering system, with detailed project information required for accurate quotations and timelines.
- Stability and Efficacy: SAMiRNA™ demonstrates high stability in circulation and effective in vivo silencing in cancer models.
- Targeted Delivery: Capable of tumor-specific delivery with minimal off-target effects.
- Toxicity and Safety: Exhibits low toxicity and cytokine induction, with no significant adverse effects observed in repeated dose studies.
- Genetic Toxicology: No mutagenic activity or chromosome aberrations detected in high-dose tests.
SAMiRNA™ represents a significant advancement in RNAi drug development, offering a safe and effective solution for gene silencing with potential applications in various diseases.
Catalog excerpts
siRNA Synthesis Service miRNA Synthesis Service In vivo siRNA Custom Service: SAMiRNA™
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siRNA Synthesis Service AccuTargetTM siRNAs ··················································································································································· 33 Genome-wide Predesigned siRNA, Primer & Control AccuTarget™ Genome-wide Predesigned siRNA Library ··············································································· 35 AccuTarget™ Real-Time PCR Primers Library ······································································································ 36 AccuTarget™ Control siRNAs ····································································································································...
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Gene knockdown technologies, such as antisense, ribozyme, and gene knockouts are used to perform loss-of-function studies. However, the post-genomics era calls for high-throughput gene function studies, at which the prior gene knockdown approach are provide due to poor reproducibility, high cost, and excessive time to the result. Thus, the advent of short interfering RNA or siRNA technology has opened up many new possibilities in the field of gene suppression. siRNA has the following advantages over other RNAi technology: • Reduced time and costs: Less screening is required to obtain highly effective...
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AccuTarget™ siRNA The most critical among these factors is the design of effective and specific siRNA. Bioneer, in collaboration with the world renowned National Genome Information Center (NGIC) at KRIBB institute, has developed Turbo si-Designer, which is a proprietary siRNA selection algorithm. Turbo si-Designer identifies highly effective siRNA target sites with exceptional success rates. Several important parameters including base composition, the number of repetitive bases in a row, thermodynamic instability, energy profiling and base preference were considered in the development of Turbo...
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AccuTarget™ Genome-wide Predesigned siRNA Library ■ Description Bioneer offers over 132,000 predesigned siRNAs for more than 44,000 target genes from Human, Mouse and Rat. Search our extensive siRNA library by Gene ID, Symbol, Synonyms, Description, or Accession Number. Once you find your gene of interest, choose your guaranteed yield and purification level, and check your predicted siRNA knockdown efficiency. You can even order your qPCR primers for siRNA knockdown validation. Convenient and easy siRNA ordering - only from Bioneer. AccuTarget™ Genome-wide Predesigned siRNA Library can be used...
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■ Features and Benefits • In ready-to-ship format: 11,154 human genes specific primers • All Primers pre-validated • Competitive pricing: Great value for your research dollar ■ Description AccuTarget™ Human Validated Real-Time PCR Primer Library Set is comprised of highly specific and sensitive Real-Time PCR primer Sets that are bioinformatically designed and validated based on the human genome. The Real-Time PCR Primer Library consists of 11,154 primer Sets validated in Real-Time PCR with SYBR Green detection using This AccuTarget™ Human Validated Real-Time PCR Primer Library Set is categorized...
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