What are the common misconceptions in siRNA synthesis?

SiRNA synthesis is a method for synthesizing small interfering RNA molecules targeting specific genes, commonly used for gene silencing and functional research. However, in the process of siRNA synthesis, there are often some misunderstandings that need to be paid attention to and avoided. The following are common misconceptions in siRNA synthesis:

1. siRNA sequence design error: The design of siRNA is the most critical step in siRNA synthesis. The selection of siRNA sequences should follow certain rules, including length, GC content, pairing stability, etc. If the siRNA sequence design is incorrect, it may result in siRNA not being able to bind to the target mRNA properly, thereby affecting the silencing effect of siRNA.

2. Poor purity: If purity control is not taken into account during siRNA synthesis, impurities may be present in siRNA, affecting its stability and effectiveness. Therefore, in the process of siRNA synthesis, it is necessary to ensure that the obtained siRNA is of high purity.

3. Unstable structure: siRNA is a double stranded RNA with a complex structure that is easily degraded by nucleases within cells. If the structure of siRNA is unstable, it may lead to siRNA degradation and fail to exert its silencing effect. Therefore, in the process of siRNA synthesis, it is necessary to ensure the stability of siRNA structure.

4. High or low concentration: The concentration of siRNA has a significant impact on its silencing effect in cells. If the concentration of siRNA is too high, it may lead to non-specific side effects within cells; If the concentration of siRNA is too low, it may lead to a less significant silencing effect of siRNA. Therefore, strict control of the concentration of siRNA is required during the synthesis process.

5. Improper targeting selection: Targeting selection of siRNA is one of the key steps in siRNA synthesis. If the selected siRNA targeting is not accurate, it may result in the siRNA not achieving the expected silencing effect. Therefore, careful consideration needs to be given to the targeting selection of siRNA during the synthesis process.

Overall, siRNA synthesis is a complex task that requires careful attention at every step. Only by ensuring the accurate design, high purity, stable structure, appropriate concentration, and correct targeting selection of siRNA can efficient siRNA be obtained and play its role in gene silencing and functional research. I hope the common misconceptions in siRNA synthesis described in this article can be helpful to readers, raise their awareness, and avoid mistakes.