Fluorescently Labeled siRNA

As a professional provider of biochemical services, BOC Sciences can offer a variety of Fluorescently labeled siRNA services. With years of expertise in labeling technology, we offer customized solutions, including FITC-labeled siRNA, Cy5-labeled RNA, Cy3-labeled RNA, and more, to meet your specific project requirements.

What is Fluorescently Labeled siRNA?

Fluorescently labeled siRNA is a technique in which fluorescent dyes are added to siRNA molecules through a chemical reaction. This labeling method allows siRNA molecules to be detected under fluorescence microscopy, allowing researchers to monitor the behavior and action of siRNAs in real-time. Fluorescently labeled siRNAs track the uptake and distribution of siRNAs in cells or animals, and the transfection efficiency of siRNAs in cells can be observed using fluorescence microscopy and flow cytometry, and the localization of siRNAs can be observed using laser confocal microscopy. It is usually labeled at the 5' end, and common fluorescent markers are FAM, Cy3, and Cy5.

Mechanism of Fluorescently Labeled siRNA

Fluorescently labeled siRNA technology is a method of attaching fluorescent dyes to specific sites of siRNA through covalent bonds. This labeling does not significantly affect the biological function of the siRNA, allowing researchers to monitor the siRNA in real-time without altering its ability to target. As the fluorescently labeled siRNA enters the cell and fluoresces under a specific wavelength of excitation light, it is easy to detect and analyze by fluorescence microscopy or flow cytometry.

Application of Fluorescently Labeled siRNA

Fluorescently labeled siRNA has a wide range of applications in a number of research fields, the following are its main application areas and functions:

Measure siRNA uptake efficiency

The cell absorption process of siRNA can be monitored in real-time by fluorescent labeling. This approach not only assesses the efficiency of siRNA uptake in different cell types, but also enables comparison of the effects of different delivery systems. With this approach, the researchers were able to optimize siRNA delivery vectors and select the most effective delivery strategies to improve siRNA uptake and function.

Optimize transfection experiments

Fluorescently labeled siRNAs can be used to analyze siRNA stability and transfection efficiency. Labeled siRNAs can also be used as siRNA intracellular localization and dual-labeling assays (with labeled antibodies) to track cells that have introduced siRNAs during transfection, combining transfection with downregulation of target protein expression.

Analyze the distribution of siRNAs within cells

Fluorescent labeling enabled the researchers to locate siRNA in the cytoplasm and nucleus. This is crucial for understanding the mechanism of action of siRNA. For example, by looking at the distribution of siRNA within cells, it is possible to infer where they interact with target mRNA, leading to a better understanding of the details of the RNAi silencing process.

Assess the stability of the siRNA

By monitoring changes in the fluorescence intensity of fluorescently labeled siRNA within cells, researchers can assess the rate of degradation of siRNA. This is important for the development of more stable siRNA molecules, which could help design siRNA with a longer half-life, thereby improving its efficacy in therapy.

Verify the silencing effect of the target gene

With colocalization analysis, researchers can assess the silencing efficiency of RNAi. This method colocalizes the fluorescently labeled siRNA with the target gene mRNA to see if the siRNA successfully binds and silences the target gene. This not only verifies the targeting of siRNAs, but also quantifies their silencing efficiency, thus providing data support for siRNA design and optimization.

Examples of Fluorescently Labeled siRNA

FITC-labeled siRNA

Fluorescein isothiocyanate (FITC) is a green fluorescent dye widely used in cell and molecular biology experiments. The researchers injected the FITC-labeled siRNA into the tumor, and fluorescence microscopy can track the path and location of the siRNA once it enters the cell in real-time

Cy3-labeled siRNA

In neuroscience studies, researchers use Cy3-labeled siRNA to inhibit the expression of specific genes, while using fluorescence microscopy to analyze the distribution and change of siRNA in cells to understand the effect of gene silencing on cells.

Fig.1 DAPI staining of Cy3-siRNA uptake in rat myocardium. (Wang Leo L., et al., 2017)

Cy5-labeled siRNA

In immunology studies, the use of Cy5-labeled siRNA can show its uptake and distribution in different immune cell types, and the distribution and uptake of siRNA can be analyzed by flow cytometry.

Alexa Fluor 488-labeled siRNA

The Alexa Fluor family includes fluorescent dyes in a variety of colors, such as Alexa Fluor 488 (green) and Alexa Fluor 647 (red), with high light stability and high fluorescence intensity. In cancer research, Alexa Fluor 488-labeled siRNAs were used to inhibit the expression of tumor-associated genes, and the localization and persistence of siRNAs in tumor cells were observed by live-cell imaging technology. Used in cancer research to assess siRNA stability and gene silencing efficiency.

Advantages of Fluorescently Labeled siRNA

Fluorescent-labeled siRNA has the following advantages:

Real-time monitoring: It can observe the dynamic changes of siRNA in cells in real-time, helping researchers to understand the uptake and transportation of siRNA.

High sensitivity: The high sensitivity of fluorescent labeling facilitates the detection of low concentrations of siRNA, and its location and dynamics can be clearly seen even in complex cellular environments.

Quantification: Quantitative analysis of data by fluorescence intensity can help quantify siRNA and ensure reliable and repeatable experimental results.

Multi-purpose: Fluorescent labels of different colors can be used for multi-target analysis at the same time, enabling researchers to detect the interaction and action of multiple targets in the same experiment, increasing the flexibility and information of the experiment.

Our Fluorescently Labeled siRNA services

BOC Sciences offers a comprehensive range of Fluorescently labeled siRNA services, from design to synthesis to labeling. Features of our services include:

Customized design: Design the most suitable siRNA sequence and labeling strategy according to the specific needs of the customer.

High-quality synthesis: Advanced synthesis technology is used to ensure high purity and high performance of siRNAs.

Multiple labeling options: A variety of fluorescent labeling options, including FITC, Cy5, Cy3, etc., are available to meet different experimental needs.

Professional technical support: Provide comprehensive technical support and consulting services to help customers solve various problems in experiments.

With our professional services, you can get high-quality Fluorescently labeled siRNA that will help you succeed in your research project.

FAQ

1. What is fluorescently labeled siRNA?

Fluorescently labeled siRNAs are fluorescent dyes attached to small interfering RNA (siRNA) molecules so that they can be detected and tracked in cells or tissues to study the distribution and mechanism of action of siRNAs.

2. What are the commonly used dyes for fluorescently labeled siRNAs?

Commonly used fluorescent dyes include FITC (fluorescein isothiocyanate), Cy3, Cy5, and Alexa Fluor dyes. These dyes have different emission spectra and can be used for multiplexing and multicolor analysis.

3. Does fluorescent labeling affect the function of siRNAs?

Fluorescent labeling may affect the function of the siRNA, depending on the type of dye and the location of the label. In general, labeling at the 5' or 3' end of the siRNA reduces interference with its gene silencing activity.

4. How do I verify the success of fluorescently labeled siRNAs?

Fluorescence microscopy can be used to observe the fluorescence signal, HPLC, or mass spectrometry to detect marker binding, and functional experiments to verify that the labeled siRNA is still effective in silencing the target gene.

5. How stable are fluorescently labeled siRNAs in vivo?

The stability of fluorescently labeled siRNAs in vivo depends on the siRNA itself and the chemistry of the fluorochrome. Protective modifications, such as methylation modifications, can improve the stability of siRNAs and reduce degradation in vivo.