Rnai Mechanism
Rnai Mechanism - Related Videos
09:51
RNAi in C. elegans
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2023
RNA interference (RNAi) is a widely used technique in which double stranded RNA is exogenously introduced into an organism, causing knockdown of a target gene. In the nematode, C. elegans, RNAi is particularly easy and effective because it can be delivered simply by feeding the worms bacteria that express double stranded RNA (dsRNA) that is complementary to a gene of interest. First, this video will introduce the concept of RNA interference and explain how it causes targeted gene knockdown.
07:56
Bacterial Delivery of RNAi Effectors: Transkingdom RNAi
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Cited by 18 •
2010
For development of RNA interference (RNAi)-based therapies, a novel strategy was developed, transkingdom RNAi (tkRNAi). This technology uses non-pathogenic bacteria to produce and deliver therapeutic short hairpin RNA (shRNA) into target cells. Here, tkRNAi was successfully applied for reversal of classical ABCB1-mediated multidrug resistance (MDR) of cancer cells.
02:29
Culturing and Maintenance of RNAi-Expressing Bacteria
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2025
Source: Anderson, Q. L., et al., A High-throughput, High-content, Liquid-based C. elegans Pathosystem. J. Vis. Exp. (2018)This video demonstrates the preparation of RNA interference (RNAi) bacteria carrying plasmids to inhibit target gene expression. It outlines RNAi bacterial selection, growth, and induction steps to produce dsRNA-expressing plates for downstream assays.
09:40
RNAi Screening to Identify Postembryonic Phenotypes in C. elegans
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Cited by 9 •
2012
We describe a sensitized method to identify postembryonic regulators of protein expression and localization in C. elegans using an RNAi-based genomic screen and an integrated transgene that expresses a functional, fluorescently tagged protein.
15:31
MISSION esiRNA for RNAi Screening in Mammalian Cells
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Cited by 17 •
2010
Here we use a human esiRNA library in a high-throughput screen for genes involved in cell division. We demonstrate how to set up and conduct an esiRNA screens, as well as how to analyze and validate the results.
FAQs
Dicer converts double-stranded RNA into short interfering RNAs, creating the small-RNA signals needed for targeting. One strand then loads into the RNA-induced silencing complex, or RISC. This assembly connects RNA processing with gene regulation because the retained guide strand provides the sequence information used to recognize a matching messenger RNA.
The guide strand directs RISC toward messenger RNA containing a complementary sequence. This sequence recognition gives RNAi its specificity, allowing regulation of a particular gene transcript rather than producing a general reduction in protein production. Once the target is recognized, Argonaute can promote messenger RNA cleavage or contribute to translational repression.
Targeted silencing can occur through more than one downstream effect. In the canonical pathway, Argonaute may cleave a complementary messenger RNA, reducing the transcript available for protein production. Alternatively, the matched transcript may undergo translational repression, which limits protein synthesis without the same cleavage outcome. These mechanisms provide distinct routes to reduced gene expression.
Researchers use RNAi to reduce production of a selected protein and then examine the resulting biological effects. This creates a targeted loss-of-function approach, linking a gene or its protein product to a cellular or organismal process. Comparing silenced conditions with appropriate nonsilenced conditions can help reveal the function of endogenous genes.
Reducing expression of a selected gene or protein allows investigators to examine how that factor contributes to a disease pathway. The resulting changes can help identify relationships between gene activity and disease-related processes. Because the mechanism is sequence-specific, RNAi supports focused interrogation of particular molecular targets rather than relying only on broad changes in gene expression.
RNAi provides a model for deliberately lowering production of specific proteins through sequence-guided silencing. This principle is relevant to therapeutic research because disease-associated pathways may depend on particular gene products whose reduction could alter the pathway. The same mechanism also reflects biological defenses against some foreign genetic material, connecting basic biology with treatment development.