What can siRNA be used for?

siRNA is a synthetic RNA duplex designed to specifically target a particular mRNA for degradation. siRNAs can also be used for knockdown of non-protein coding genes, such as long noncoding RNAs (lncRNA).

What produces siRNA?

In this method, long dsRNAs are prepared by in vitro transcription using a template that typically encodes a 200–1000 nt region of the target mRNA. The dsRNA is then digested in vitro with RNase III (or Dicer) to produce a population, or cocktail, of siRNAs.

How much does siRNA cost?

Standard siRNA Synthesis

Are siRNA biologics?

siRNA is the molecule that carries out the process of RNAi. siRNA is distinct from other types of biologics, such as antibodies, which bind to already-formed proteins and impact their function or remove them from the body.

Does siRNA occur naturally?

Structure. Naturally occurring siRNAs have a well-defined structure that is a short (usually 20 to 24-bp) double-stranded RNA (dsRNA) with phosphorylated 5′ ends and hydroxylated 3′ ends with two overhanging nucleotides. The Dicer enzyme catalyzes production of siRNAs from long dsRNAs and small hairpin RNAs.

How does siRNA knockdown work?

Gene knockdown by this method is achieved by introducing small double-stranded interfering RNAs (siRNA) into the cytoplasm. Small interfering RNAs can originate from inside the cell or can be exogenously introduced into the cell. After the RISC localizes to the target mRNA, the RNA is cleaved by a ribonuclease.

What is the first step in the production of a siRNA?

What is the first step in the production of a miRNA? Complementary base pairing allows the miRNA or siRNA and the mRNA to interact.

How do you dissolve siRNA?

General considerations: Dissolve siRNA in sterile water and prepare 5 µl aliquots, store at –80°C. For use, dilute siRNA in siRNA dilution buffer (diluted siRNA is not stable, use only once). 5x siRNA buffer aliquots (Dharmacon) are stored at 4°C in the cell culture lab.

How does siRNA affect gene expression?

The siRNA-induced post transcriptional gene silencing starts with the assembly of the RNA-induced silencing complex (RISC). The complex silences certain gene expression by cleaving the mRNA molecules coding the target genes. This cleavage results in mRNA fragments that are further degraded by cellular exonucleases.

How do you get siRNA into cells?

After entering the tissue interstitium, siRNA is transported across the interstitial space to the target cells. After reaching the target cell, siRNA undergoes internalization via endocytosis, a process that involves siRNA being encapsulated in endocytic vesicles that fuse with endosomes.

Is siRNA the same as RNAi?

The major difference between siRNAs and miRNAs is that the former inhibit the expression of one specific target mRNA while the latter regulate the expression of multiple mRNAs. A considerable body of literature now classifies miRNAs as RNAi molecules.