siRNA Labeling

BOC Sciences is a leading provider of siRNA Labeling services. With years of expertise in nucleic acid labeling technology, we offer customized solutions to meet your specific project requirements. Our dedicated scientific team uses first-hand knowledge and state-of-the-art technology to ensure superior results in labeling applications.

What is siRNA Labeling?

siRNA (small interfering RNA) labeling refers to the attachment of various tags (such as fluorophores, biotin, enzyme labels, etc.) to siRNA molecules by chemical or biological means. This labeling method allows researchers to track and analyze siRNA distribution, stability, uptake efficiency, and targeted gene silencing in and out of the cell. siRNA labeling technology has important application value in RNA interference (RNAi) research.

Methods of siRNA Labeling

Fluorescent labeling

Commonly used fluorescent dyes include FITC, Cy3, Cy5, and Alexa Fluor, which can be used by fluorescence microscopy or flow cytometry to track and quantify siRNA distribution, transduction efficiency, and targeting effects in real time. Fluorescently labeled siRNAs facilitate the study of intracellular dynamics of siRNAs and their interactions with target mRNAs.

Biotin labeling

Biotin labeling is a highly sensitive and specific labeling technique. Detection and capture of siRNAs can be achieved by covalently attaching biotin molecules to siRNAs and then binding to streptavidin to form highly stable complexes. Biotinylated siRNAs are commonly used in experiments such as immunoprecipitation, affinity purification, and ELISA.

Enzyme labeling

Enzyme labeling is the conjugation of enzymes (such as horseradish peroxidase, HRP) to siRNAs to achieve detection through chemiluminescence or chromogenic reactions. This method is simple to operate and has a significant signal amplification effect, and is often used in Western blot, immunohistochemistry, and other experiments.

Radioisotope labeling

Radiolabeling refers to the introduction of radioisotopes (such as ³²P, ¹²⁵I, etc.) into siRNA molecules in order to detect the presence and distribution of siRNAs by autoradiography or counters. This method is highly sensitive, but due to the dangerous nature of the use of radioactive materials, special caution is required in experimental handling and waste disposal.

Fig.1 Strategies of HEN1-directed labeling of small RNAs. (Plotnikova, Alexandra, et al., 2014)

Mechanism of siRNA Labeling

The mechanism of action of siRNA labeling includes the following:

• Detect siRNA uptake: By labeling siRNAs, it is possible to track their efficiency and pathways into cells in real-time. This labeling, which typically uses fluorescent dyes or radioisotopes, allows researchers to observe and document how the siRNA is taken up by cells under a microscope, optimizing the siRNA delivery system and improving transfection efficiency.
• Analyze siRNA distribution: Labeled siRNAs can help study their distribution within cells. Using fluorescence microscopy or other imaging techniques, researchers can precisely determine the localization of siRNAs in the cytoplasm and nucleus, understanding their transport and localization processes within the cell.
• Assess siRNA stability: With labeling, researchers can monitor the rate and stability of siRNA degradation within cells. The change of fluorescence intensity or radioactive signal of labeled material over time can reflect the stability of siRNA in the cell.
• Verify the silencing effect of the target gene: The silencing efficiency of RNAi can be assessed by combining the tagged siRNA with the colocalization analysis of the target gene mRNA. Fluorescence colocalization allows researchers to observe the binding of the siRNA to the target mRNA in the cell, thereby verifying the silencing effect of the siRNA on the target gene.

Advantages of siRNA Labeling

• Real-time tracking: Fluorescent labeling allows for real-time monitoring of intracellular siRNA dynamics. Fluorescent labeling allows researchers to observe the localization and movement of siRNAs within cells to understand their role and mechanisms in cellular processes. This real-time tracking technology is essential for studying the function and mechanism of action of siRNAs.
• Diversity: A variety of labeling methods and label types are available to meet the needs of different studies. Researchers can choose the labeling method that suits their experimental conditions and goals, such as fluorescent, radioisotope, and enzyme labeling.
• Precise quantification: siRNAs can be precisely quantified by radioisotope or enzyme labeling. Radioisotope labeling provides highly sensitive quantitative detection for experiments that require extreme precision. Enzymatic labeling, on the other hand, can be quantified by producing a detectable signal through an enzymatic reaction.

Applications of siRNA Labeling

Intracellular localization studies

siRNA labeling technology enables researchers to observe the distribution and transduction efficiency of siRNAs within cells in real-time. Through fluorescence microscopy, the localization of siRNA in the cell can be visualized, which can help optimize the delivery system and method of siRNA and improve the efficiency of gene silencing.

Targeted gene silencing

siRNA labeling can help validate the targeting effect of siRNAs. By detecting the distribution of the labeled siRNA in a cell or tissue, it can be determined whether it has successfully reached the target region and exerts gene silencing. This is essential for preclinical research and evaluation of siRNA therapeutics.

Research on drug delivery systems

Labeled siRNAs can be used to assess the transduction efficiency and in vivo distribution of different drug delivery systems, such as lipid nanoparticles, polymer-carriers, etc. By comparing the behavior of labeled siRNAs in different delivery systems, the optimal delivery vector can be screened to improve the therapeutic effect of siRNAs.

Molecular interaction studies

siRNA labeling technology can be used to study the interaction of siRNAs with their target mRNA, proteins, and other biomolecules. Through techniques such as co-immunoprecipitation, these interacting complexes can be captured and analyzed, providing insight into siRNA-mediated gene silencing mechanisms.

Our siRNA Labeling Services

We offer a comprehensive range of siRNA labeling services, including design, synthesis, and characterization, to ensure high-quality labeled siRNAs to meet your research needs. Our services include:

• Customized design: Design sequence-specific siRNAs according to customer needs and select appropriate tags for labeling.
• Efficient synthesis: siRNA products that use advanced synthesis techniques to ensure high purity and high labeling efficiency.
• Quality Control: Characterize and validate labeled siRNAs with a variety of assays to ensure their quality and functionality.
• Technical support: Provide professional technical support and consulting services to help customers solve problems in experiments.
• With our siRNA labeling services, you can get high-quality labeled siRNAs that can help you achieve breakthroughs in your research.

FAQ

1. What is siRNA labeling?

siRNA labeling refers to the attaching of fluorescent dyes, radioisotopes, or other detectable tags to small interfering RNA (siRNA) molecules in order to track their location and behavior within cells or in vivo, and to study their biodistribution and mechanism of action.

2. What are the commonly used siRNA labeling methods?

Commonly used siRNA labeling methods include fluorescent dye labeling (e.g., FITC, Cy3, Cy5), radioisotope labeling, and biotin labeling. These tags can be attached to siRNAs by chemical conjugation or enzymatic reactions.

3. Does siRNA labeling affect its function?

Labeling of siRNAs may affect their function, depending on the location and type of label. In general, avoiding labeling in functional regions of the siRNA (e.g., 5' or 3' ends) can reduce the impact on siRNA silencing efficiency.

4. How can I verify the success of siRNA labeling?

The success of siRNA labeling can be verified by a variety of methods, such as fluorescence microscopy to observe the presence of labeled fluorescence, high-performance liquid chromatography (HPLC) or mass spectrometry to detect marker binding, and functional experiments to verify that the labeled siRNA is still effective in silencing the gene of interest.

5. What is the stability of siRNA labeling in vivo?

The stability of siRNA labeling depends on the chemistry of the marker and the protective measures of the siRNA. For example, fluorescently labeled siRNAs can be rapidly degraded in vivo, requiring the use of protective modifications such as 2'-O-methylation modifications to improve their stability.