What is the RNAi mechanism?
RNA interference (RNAi) is a regulatory mechanism of most eukaryotic cells that uses small double-stranded RNA (dsRNA) molecules as triggers to direct homology-dependent control of gene activity (Figure 1) (1).
Why do cells have this RNAi mechanism?
RNAi may be used for large-scale screens that systematically shut down each gene in the cell, which can help to identify the components necessary for a particular cellular process or an event such as cell division. The pathway is also used as a practical tool in biotechnology, medicine and insecticides.
How is RNAi performed in c elegans?
There are three ways to carry out RNAi in C. elegans: injection (Fire et al., 1998), soaking (Tabara et al., 1998), and feeding (Timmons and Fire, 1998). All three can produce efficient gene knockdowns.
How is the target gene silenced using RNAi mechanism?
Double-stranded RNA-mediated interference (RNAi) is a simple and rapid method of silencing gene expression in a range of organisms. The silencing of a gene is a consequence of degradation of RNA into short RNAs that activate ribonucleases to target homologous mRNA.
Why do cells have RNAi mechanism and why this mechanism is important to researchers?
Simple interfering RNA produced through the RNAi mechanism acts as an intracellular protection mechanism against invading viruses. In the laboratory, researchers utilize this method to block protein synthesis from target genes. It is simple and easy to synthesize and design a siRNA that binds to complementary mRNA.
How does RNAi get 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.
How is siRNA introduced?
siRNAs can also be introduced into cells by transfection. Since in principle any gene can be knocked down by a synthetic siRNA with a complementary sequence, siRNAs are an important tool for validating gene function and drug targeting in the post-genomic era.
Which of the following is used as a vector for gene transfer during RNAi?
As a versatile gene vector, bacteria have been employed and shown to be an effective, safe and inexpensive measure for delivering RNAi to mammalian cells.
How is RNAi used in lab?