siRNA synthesis is the simplest method to achieve gene silencing in the body or cell, with low cost and high transfection efficiency. It is also a means of RNAi clinical treatment and has irreplaceable advantages in drug development. BOC Sciences can provide customers with common chemically synthesized siRNAs, as well as a wide range of modified siRNAs depending on customer requirements.
When the most effective siRNA has been found, it is suitable to use nucleotide monomers as raw materials to chemically synthesize two complementary RNA single strands with a length of about 21-23 nt, and then anneal to form a double-stranded complex. The obtained product has high purity, high interference efficiency and simple synthesis.
The in vitro transcription method uses two complementary DNA as a template, connects the T7 promoter upstream of the sense strand and antisense strand targeting the target sequence, and T7 RNA polymerase is used to transcribe each of them in vitro to obtain two single-stranded RNAs. In the in vitro transcription method, two complementary DNAs are used as templates, a T7 promoter is connected to the upstream of the sense strand and the antisense strand of the target sequence, and T7 RNA polymerase is used to transcribe each of them in vitro to obtain two single-stranded RNAs. Anneal the two single strands to form a double-stranded RNA, and then digest it with RNase. The resulting product is the desired double-stranded RNA after purification. This method is suitable for screening effective siRNAs, but it is not suitable for long-term research on specific siRNAs.
The enzymatic digestion method is to chemically synthesize a 200-1000 bp complete long fragment dsRNA of the target gene in vitro, which is then digested with the key siRNA enzyme-Dicer enzyme or RNase III formed in the cell, and various siRNA mixtures targeting different sites of the same target gene can be obtained. The mixture is then transfected into the cells to obtain a high inhibition efficiency. This method has a high inhibition rate, but the mixture produced by this method may cause non-specific inhibition. In addition, it is difficult to transcribe long-chain RNA in vitro, and the cost of Dicer enzyme is high. This method is not suitable for long-term research projects, or requires a specific siRNA for research, especially for gene therapy.
In vivo transcription is the transfer of siRNA plasmids, viral expression vectors or PCR products with siRNA expression cassettes into cells, where cellular expression produces RNA interference effects.
Fig. 1 siRNA mechanism of action (Halib N, 2022)
|Chemically modified siRNA
|Fluorescence labeled siRNAs
|Custom specific siRNA synthesis
Provide gene sequence, Gene ID or Accession Number, etc.
Validated siRNA or miRNA fragments and related reports.