* Only for research. Not suitable for any diagnostic or therapeutic use.
RNAi technology can be used to assess the function of many genes in the genome that may be related to the disease. It provides an effective means of blocking the expression of specific genes and evaluating their chances of compounds or signaling pathways. Especially compared to other nucleotide-based methods (such as antisense or ribozymes), RNAi is a specific, effective, and very successful method of research on the loss of function.
The laboratory application of RNAi phenomenon and gene silencing induced by siRNA in mammalian cells provide a powerful biological research tool for identifying the function and expression pathway of specific genes. siRNA gene silencing has the therapeutic potential to treat many diseases.
Successful, effective RNAi experiments depend on the efficient delivery of functional (and undegraded) siRNA into cells. Therefore, choosing the most suitable transfection reagent and optimizing the experimental protocol is crucial to the success of the experiment.
The most common delivery methods are cationic and virus-mediated transfection.
Lipid transfection Cationic lipids with long hydrophobic chains with long positively charged head groups interact with negatively charged siRNAs, wrap them in lipid bilayers, and then endocytose them.
Cationic polymer-based nanoparticles Reduce cytotoxicity, increase transfection efficiency, and allow the delivery of modified siRNA.
In electroporation, electrical pulses redirect phospholipids, forming pores in the membrane, allowing siRNA to enter the cell. Electroporation is usually used for cells that are difficult to transfect. However, specific settings (voltage, pulse number, and pulse length) must be optimized for each cell type.
Viral delivery of RNAi and siRNA is a powerful alternative to lipofection. Viral transduction is the best choice in almost all mammalian cell types, including difficult to transfect, primary, and even non-dividing cell type diseases. Transduction by lentivirus or retrovirus can stably integrate siRNA and RNAi into the cell genome, thereby achieving durable expression.
BOC Sciences siRNA provides optimized transfection reagents for siRNA in vivo and in vitro. All products are produced under strictly QA and QC and have been verified by external experiments.
|In vitro transfection|
The core of RNAi requires siRNA to effectively bind and act on the corresponding mRNA. The design and synthesis of siRNA is very important first. The optimal design can achieve a satisfactory silent effect with the minimum working concentration and reduce the occurrence of side reactions.
It should be emphasized that a negative control needs to be set at the same time to exclude the non-specific silencing phenomenon and the positive control to confirm the effectiveness of the entire experimental system.
It should be noted that the synthesis of siRNA must select high-purity siRNA. The purity of siRNA is directly related to transfection efficiency and silencing efficiency. The working concentration of siRNA is related to siRNA design, cell type, and target gene.
It is recommended to carry out relevant pre-experiments to optimize each condition.
Choosing the right siRNA transfection reagent is another key to the success of RNAi experiments, to deliver at the highest efficiency, resulting in using siRNA to suppress gene expression while avoiding cytotoxicity. Effective transfection requires that the transfection reagent can effectively deliver RNAi/siRNA while being gentle on the cells. Maintaining low cytotoxicity is important because the toxicity mediated by the transfection reagent may obscure the true phenotype of the target gene under study.