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GeticoFect Exosome Transfection Reagent Instruction Manual1. IntroductionThe GeticoFect Exosome Transfection Reagent, developed by Geneticsci Biotech (Shanghai) Co., Ltd., is a rev
GeticoFect Exosome Transfection Reagent Instruction Manual1. IntroductionThe GeticoFect Exosome Transfection Reagent, developed by Geneticsci Biotech (Shanghai) Co., Ltd., is a rev
Catalog Number | Specification | Price (CNY) |
181501 | 0.3 mL | 2,500.00 |
181502 | 1 mL | 7,350.00 |
181503 | 10 mL | 58,800.00 |
• Product Name: GeticoFect® Exosome Transfection Reagent
• Storage Condition: Store at 4°C (Do not freeze)
• Manufacturer: Getico Scientific (Shanghai) Co., Ltd.
In the current cutting-edge field of life sciences research, exosomes, as key mediators of intercellular communication, show great application potential in disease diagnosis, treatment, and drug delivery. The efficient loading of specific nucleic acid molecules (such as siRNA and miRNA) into exosomes is crucial for in-depth exploration of exosome functions and expansion of their application scope. GeticoFect® Exosome Transfection Reagent is specially designed and optimized for exosome transfection. Like a precise key, it helps researchers successfully deliver siRNA, miRNA, and other molecules into various exosomes, opening a new chapter for exosome-related research.
GeticoFect® Exosome is a proprietary and highly optimized RNAi-specific nano-formulation. The ingenious application of nanotechnology endows this reagent with unique advantages. Its nanoparticles have a small size and a large specific surface area, which can tightly bind to siRNA and miRNA to form stable and efficient complexes. This complex is like an intelligent transport carrier that can protect nucleic acid molecules from external interference and degradation. At the same time, relying on its special physicochemical properties, it is easier to interact with exosomes to achieve efficient transfection.
Compared with other similar products on the market, GeticoFect® Exosome Transfection Reagent has significant advantages, standing out like a bright star among many products.
GeticoFect® Exosome Transfection Reagent exhibits excellent transfection efficiency. When introducing siRNA and miRNA into exosomes, it only requires a lower concentration of RNAi and miRNA to achieve more effective gene delivery. This is due to its unique nano-formulation design, which can deliver nucleic acid molecules into exosomes more efficiently. A lower nucleic acid concentration not only reduces experimental costs but also minimizes non-specific effects. Non-specific effects may interfere with experimental results and lead to incorrect judgments about gene functions. However, GeticoFect® Exosome Transfection Reagent controls non-specific effects to a very small range while ensuring efficient gene delivery, providing a strong guarantee for the accuracy of experimental results. In addition, this reagent also has good compatibility and is suitable for miRNA transfection, providing more convenience for researchers in miRNA-related research.
This reagent is applicable to a variety of exosomes. Whether exosomes are derived from different cell types or in different physiological states, it can achieve efficient transfection. This provides a universal transfection method for exosome transfection experiments, eliminating the need for researchers to worry about selecting suitable transfection reagents when conducting research on different types of exosomes. Whether exploring the biological functions of exosomes in basic research or developing exosomes as drug delivery carriers in clinical applications, GeticoFect® Exosome Transfection Reagent can play an important role and provide strong technical support for exosome-related research.
The simplicity of experimental operation is crucial for the efficiency of scientific research. GeticoFect® Exosome Transfection Reagent has a simple and easy-to-understand experimental operation process. Researchers only need to mix GeticoFect® Exosome Transfection Reagent with siRNA/RNA and exosomes, and after a short incubation period, add the mixture to cells, followed by subsequent incubation and experimental operations. The entire process does not require complex technologies or tedious steps, reducing the difficulty and threshold of the experiment and allowing more researchers to get started easily. This simple and rapid operation method, combined with high transfection efficiency, makes GeticoFect® Exosome Transfection Reagent an ideal choice for efficient siRNA transfection into exosomes.
The high transfection efficiency of GeticoFect® Exosome Transfection Reagent ensures that a large number of siRNA and miRNA molecules are accurately delivered into exosomes, thereby achieving a high level of gene delivery effect. By using this reagent, researchers can obtain satisfactory experimental results and clearly observe the biological functions of exosomes after loading nucleic acid molecules.
Tests on various exosomes show that this product has extremely high transfection efficiency. Moreover, we are one of the few suppliers of exosome-specific transfection reagents on the market, which fully reflects the professionalism and uniqueness of our product. Choosing GeticoFect® Exosome Transfection Reagent means choosing an efficient and professional exosome transfection solution.
A: GeticoFect Exosome Transfection Reagent is designed based on advanced nanotechnology and a unique chemical formula. Its core principle is to use special cationic polymers that can form stable and biocompatible complexes with negatively charged nucleic acids (such as DNA and RNA) or other biomolecules through electrostatic interaction. This complex can be efficiently taken up by exosomes while effectively protecting biomolecules from degradation. Compared with other exosome transfection reagents, GeticoFect Exosome has the following unique advantages:
• Higher transfection efficiency: In transfection experiments of exosomes derived from various cells, its transfection efficiency can be 30%-50% higher than that of ordinary reagents. For example, for exosomes derived from human umbilical vein endothelial cells (HUVEC), the use of GeticoFect Exosome can increase the transfection efficiency of small interfering RNA (siRNA) to more than 80%.
• Wide applicability to biomolecules: It is not only suitable for common nucleic acid substances, such as siRNA, microRNA (miRNA), messenger RNA (mRNA), and plasmid DNA, but also can effectively transfect some small molecule drugs, proteins, and other biomolecules, expanding the application range of exosomes as delivery carriers.
• Low cytotoxicity: It has low toxicity to donor cells and recipient cells, which can maximize the maintenance of cell activity and normal physiological functions. This is particularly important for experiments that require maintaining the normal state of cells, such as cell function research and in vivo transplantation experiments. When transfecting exosomes derived from primary neuron cells, the survival rate of donor neuron cells can still be maintained at about 90% after using GeticoFect Exosome Transfection Reagent.
A: GeticoFect Exosome Transfection Reagent has a wide range of applicability and is suitable for the transfection of exosomes derived from various cell types. This includes, but is not limited to, common cell lines such as exosomes derived from human embryonic kidney cells (HEK293), human cervical cancer cells (HeLa), and mouse embryonic fibroblasts (NIH/3T3). At the same time, it can also achieve good transfection effects on exosomes derived from some primary cells, such as primary hepatocytes, primary immune cells, and primary neuron cells. However, exosomes derived from different cell types may have differences in surface characteristics and membrane composition. Before conducting large-scale experiments, it is recommended to perform small-scale preliminary experiments to optimize transfection conditions to ensure the best transfection efficiency. For example, due to the complexity of surface antigens of exosomes derived from primary immune cells, it may be necessary to appropriately adjust the ratio of transfection reagent to biomolecules to improve transfection efficiency.
A: GeticoFect Exosome Transfection Reagent can efficiently transfect various types of biomolecules into exosomes, mainly including:
• Nucleic acids:
◦ Small interfering RNA (siRNA): It can be used to silence specific genes in exosome recipient cells, for studying gene functions or gene therapy-related research. For example, transfecting siRNA targeting tumor-related genes into exosomes and then delivering the exosomes to tumor cells can effectively inhibit the expression of target genes in tumor cells.
◦ MicroRNA (miRNA): miRNA plays an important regulatory role in cell growth, differentiation, apoptosis, and other processes. By transfecting miRNA into exosomes, the physiological functions of recipient cells can be regulated. For example, transfecting miRNA with anti-inflammatory effects into exosomes and then delivering them to inflammatory cells can reduce the inflammatory response.
◦ Messenger RNA (mRNA): Transfecting mRNA into exosomes can enable recipient cells to express specific proteins. For example, transfecting mRNA encoding therapeutic proteins into exosomes for research on protein replacement therapy.
◦ Plasmid DNA: It can be used to achieve stable gene expression in recipient cells. For example, transfecting plasmid DNA carrying a reporter gene (such as green fluorescent protein gene) into exosomes, and the transfection and delivery efficiency of exosomes can be evaluated by observing the fluorescent signal in recipient cells.
• Small molecule drugs: Some small molecule drugs with therapeutic potential, such as anti-cancer drugs and neuroprotective agents, can be loaded into exosomes through GeticoFect Exosome Transfection Reagent. This helps to improve the stability of small molecule drugs, reduce their toxic and side effects, and achieve targeted delivery. For example, loading anti-cancer drugs such as doxorubicin into exosomes can enhance the targeting of drugs to tumor cells and reduce damage to normal cells.
• Proteins: Certain functional proteins, such as enzymes and antibody fragments, can also be transfected into exosomes with the help of this reagent. For example, transfecting cytokine proteins with immunomodulatory functions into exosomes can be used to regulate the immune response of recipient cells.
A: Low transfection efficiency may be caused by a variety of reasons, as follows:
• Poor exosome quality: Ensure the purity and integrity of exosomes. If exosomes are contaminated during extraction (such as other cell debris, protein aggregates, etc.) or degraded, the transfection efficiency will be seriously affected. Adopt high-quality exosome extraction methods, such as ultracentrifugation and density gradient centrifugation, and use techniques such as nanoparticle tracking analysis (NTA) and Western blot to detect the quality of extracted exosomes. For long-term storage of exosomes, they should be stored at -80°C to avoid repeated freezing and thawing.
• Problems with biomolecule quality: Ensure the quality of biomolecules to be transfected (such as nucleic acids, small molecule drugs, proteins, etc.). Nucleic acids should have high purity and be free of impurities such as proteins, RNases, and DNases. When purchasing nucleic acids, choose reputable suppliers and request quality inspection reports. For small molecule drugs and proteins, ensure their activity and stability. For example, proteins should be prevented from denaturation, and their quality can be evaluated through protein quantification and activity detection methods.
• Unoptimized transfection conditions: Conduct preliminary experiments to optimize the ratio of transfection reagent to biomolecules. Different types of biomolecules and exosomes may require different optimal transfection ratios. Generally, different ratios (μL transfection reagent: μg biomolecule) such as 1:1, 2:1, and 3:1 can be tried. Strictly follow the instructions to control the incubation time and temperature of the transfection complex. Usually, after mixing the transfection reagent with biomolecules, it is necessary to incubate at room temperature for 15-30 minutes to form a stable transfection complex.
• Influence of cell state and exosome source: Use cells in the logarithmic growth phase with good viability to extract exosomes. Excessive cell passage times, too high or too low cell density may affect the quality and transfection efficiency of exosomes. For adherent cells, the cell confluency should be controlled at 60%-80% when extracting exosomes; for suspension cells, the cell density should be adjusted to an appropriate range (such as 1×10^6 - 5×10^6 cells/mL). Regularly resuscitate cells and avoid using cells with excessive passage times. For example, for HEK293 cells, it is recommended that the number of passages does not exceed 30. At the same time, exosomes derived from different cell types may have different responses to transfection, and targeted optimization can be carried out according to specific cell types.
A: The instability of transfection efficiency and poor repeatability can be solved from the following aspects:
• Strictly control experimental operations: Ensure that the operation steps are consistent in each experiment, including the exosome extraction method, the amount of transfection reagent and biomolecule added, and the mixing method. Use high-precision pipettes and calibrate them regularly. Record each parameter in the experimental process in detail, such as exosome concentration, the ratio of transfection reagent to biomolecule, and incubation time, to facilitate subsequent analysis and adjustment. For example, when adding transfection reagents and biomolecules, add them drop by drop and mix gently to avoid violent oscillation.
• Stabilize exosome source and quality: Use cells of the same batch and stable state to extract exosomes. Establish a standardized cell culture system to ensure the consistency of cell culture conditions, including medium formula, serum batch, and medium change time. Regularly detect cells for mycoplasma to prevent cells from being contaminated by mycoplasma, which may affect exosome quality. Each time exosomes are extracted, they are obtained from cells in the same state and extracted and purified under the same conditions.
• Optimize and fix transfection conditions: Through multiple preliminary experiments, determine the optimal transfection conditions for specific exosome sources and biomolecules, and strictly follow them in subsequent experiments. For some exosomes that are sensitive to transfection conditions, different pretreatment methods (such as surface modification of exosomes) can be tried and used in combination with GeticoFect Exosome Transfection Reagent to improve the stability of transfection efficiency. For example, for exosomes derived from some primary cells, gentle treatment with specific surfactants before transfection can significantly improve the repeatability of transfection efficiency.
A: The reasons for high cytotoxicity and the solutions are as follows:
• Excessive concentration of transfection reagent: Appropriately reduce the amount of transfection reagent. Through preliminary experiments, determine the concentration of transfection reagent that can not only ensure high transfection efficiency but also keep cytotoxicity within an acceptable range. For example, start from the recommended concentration and gradually reduce the amount of transfection reagent according to a certain gradient (such as 20%), and observe the changes in cytotoxicity and transfection efficiency.
• Too long incubation time of transfection complex: Incubate strictly according to the time specified in the instruction manual. The incubation time of GeticoFect Exosome Transfection Reagent mixed with biomolecules should generally not exceed 30 minutes. Too long incubation time may increase the toxicity of the transfection complex to cells.
• Cell sensitivity to transfection reagent: For some cell lines sensitive to transfection reagents, cells can be pretreated before transfection. For example, treat cells with cell protectants (such as the antioxidant N-acetylcysteine) to improve the tolerance of cells to transfection reagents. Alternatively, adjust the cell culture conditions, such as increasing the concentration of serum in the medium, to alleviate cytotoxicity to a certain extent.
• Influence of biomolecules themselves: Some biomolecules (such as some cytotoxic small molecule drugs, interfering RNA, etc.) may be toxic to cells at high concentrations. Optimize the transfection dose of biomolecules and try to reduce their concentration while ensuring the experimental effect. At the same time, biomolecules can be modified or encapsulated to reduce their direct toxicity. For example, small molecule drugs can be encapsulated in liposomes before transfection into exosomes, which can reduce toxicity to cells.
A: Precipitation in the transfection complex may be caused by the following reasons and solutions:
• Uneven mixing of transfection reagent and biomolecule: Fully and gently mix the transfection reagent and biomolecule when mixing them. You can use a pipette to gently blow 10-15 times, or invert the EP tube to mix the two to ensure that the transfection reagent and biomolecule are fully combined. Avoid violent oscillation to prevent damage to the structure of the transfection complex.
• Excessively high concentration of transfection reagent or biomolecule: Check whether the concentration of the transfection reagent and biomolecule is too high. If the concentration is too high, it may lead to precipitation during the formation of the transfection complex. Appropriately dilute the transfection reagent and biomolecule according to the recommended concentration range. For example, for nucleic acids with excessively high concentration, they can be diluted with nuclease-free water or a dedicated nucleic acid dilution buffer.
• Inappropriate pH value or ionic strength of the solution: Ensure that the pH value and ionic strength of the solution are within the appropriate range during the preparation of the transfection complex. Generally, the transfection reagent and biomolecule should be dissolved in a buffer with a pH value of 7.2-7.4, and solutions with high concentrations of salt ions should be avoided. If the buffer used is inappropriate, it can be replaced with a recommended buffer, such as Opti-MEM, a low-serum medium.
A: This reagent should be stored in a 4°C refrigerator and should not be frozen. Freezing will damage the nanostructure and polymer components in the transfection reagent, resulting in a decrease in its transfection performance. If the reagent is accidentally frozen, after thawing, observe whether the reagent has abnormal phenomena such as stratification and turbidity. If there is an abnormality, it is not recommended to use it continuously. Under proper storage conditions, the validity period of the reagent is [X] months. When approaching the validity period, it is recommended to conduct a preliminary experiment to detect whether the transfection efficiency of the reagent has decreased. If the transfection efficiency is significantly reduced, a new reagent should be replaced in time.
A: The following precautions should be taken during the operation:
• Avoid nuclease contamination: The entire operation process should be carried out in a nuclease-free environment. Use nuclease-free pipette tips, centrifuge tubes, buffers, and other consumables. Operators should wear gloves to avoid nuclease contamination of reagents and biomolecules from hands. For example, when opening reagent bottles and biomolecule tubes, minimize the contact time with the external environment.
• Accurately measure reagents and biomolecules: Use a high-precision pipette to accurately measure the transfection reagent and biomolecule. The range of the pipette should be selected appropriately to avoid inaccurate measurement due to too large or too small a range. Before measurement, fully mix the reagent and biomolecule and let them stand at room temperature for a while to make their temperature consistent with room temperature and reduce errors.
• Follow the transfection steps: Strictly follow the steps in the instruction manual for transfection operations. This includes the preparation sequence of the transfection complex, incubation time, and the method of adding to exosomes. For example, the transfection reagent should first be diluted in serum-free medium, then the biomolecule should be added, mixed gently, and incubated. When adding the transfection complex to exosomes, add it drop by drop and gently shake the centrifuge tube to ensure uniform distribution of the complex.
• Avoid excessive handling of exosomes: Before and after transfection, minimize unnecessary manipulation of exosomes to prevent excessive damage to them. For example, during the isolation and purification of exosomes and the addition of transfection complexes, operate gently to avoid mechanical damage to exosomes. After transfection, do not frequently test or handle exosomes, as this may affect their stability and function.
