Rnai Feeding Assay
Rnai Feeding Assay - Related Videos
03:15
RNAi Feeding in Liquid Culture: A High-Throughput Method to Knockdown Gene Expression in C. elegans
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2023
RNAi is powerful genetic tool available to C. elegans researchers. This video introduces a method to treat more worms with RNAi than plate feeding by utilizing liquid culture conditions.
03:24
RNAi Plating for C. elegans Feeding: A Technique to Induce Target dsRNA Expression in E. coli
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2023
This video describes the principles behind RNAi treatment in C. elegans and demonstrates a protocol to knockdown lin-35 in a transgenic worm strain.
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.
11:48
Protocol for RNAi Assays in Adult Mosquitoes (A. gambiae)
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Cited by 21 •
2007
Reverse genetic approaches have proven extremely useful for determining which genes underly resistance to vector pathogens in mosquitoes. This video protocol illustrates a method used by the Dimopoulos lab to inject dsRNA into Anopheles gambiae mosquitoes, which harbor the malaria parasite. The technique manipulating the injection setup and injecting dsRNA into the thorax is illustrated.
FAQs
After ingestion, bacterial dsRNA becomes the starting signal for sequence-specific silencing. In Caenorhabditis elegans, cellular processing converts the dsRNA into small interfering RNAs, which provide the sequence information needed to recognize complementary messenger RNA. This targeting step connects the consumed bacterial product to a particular gene, allowing researchers to examine consequences of reducing that gene’s expression.
The targeted messenger RNA can be affected in two related ways: the RNA-induced silencing complex may promote its degradation, or it may block translation, preventing the message from producing its protein product. Because these outcomes reduce gene expression rather than directly adding a new activity, changes in immunity, infection, or disease progression can indicate the gene’s functional contribution.
Bacteria engineered to produce dsRNA against different selected genes allow researchers to examine gene function through corresponding phenotypic changes. Applying this strategy across multiple targets creates a functional comparison of genes involved in host defense, infection, or disease progression. The approach is therefore useful for identifying factors whose reduced expression produces biologically informative outcomes.
An assay begins by selecting a gene of interest and preparing bacteria engineered to produce its corresponding dsRNA. The organism then consumes those bacteria, after which researchers assess a relevant phenotype. In infection studies, the phenotype can relate to host defense or disease progression. This workflow links gene-specific exposure to an observable biological outcome.
The essential components are an organism suitable for feeding, bacteria engineered to produce dsRNA, and a selected host or pathogen gene whose expression will be reduced. The assay also requires a measurable phenotype so that the effect of knockdown can be evaluated. Together, these components connect bacterial dsRNA production, organismal consumption, gene-expression reduction, and functional interpretation.
RNAi feeding assays are especially useful when researchers need to examine many genes in a host or infection-related system. In immunology and infection, they can reveal host factors that influence defense as well as pathogen genes associated with infection or disease progression. The resulting phenotypes support functional genomics and help prioritize genes for more detailed follow-up studies.