C Elegans Rnai Screening
C Elegans Rnai Screening - 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.
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RNAi Screening to Identify Postembryonic Phenotypes in C. elegans
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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.
10:58
Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans
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We describe a protocol using C. elegans and RNAi feeding libraries that allows automated measurement of multiple parameters such as fluorescence, size and opacity of individual worms in a population. We give one example of a screen to identify genes involved in anti-fungal innate immunity in C. elegans.
15:31
MISSION esiRNA for RNAi Screening in Mammalian Cells
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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.
03:24
RNAi Plating for C. elegans Feeding: A Technique to Induce Target dsRNA Expression in E. coli
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This video describes the principles behind RNAi treatment in C. elegans and demonstrates a protocol to knockdown lin-35 in a transgenic worm strain.
FAQs
Specificity comes from sequence matching between the small interfering RNAs and messenger RNA. After delivered double-stranded RNA is processed, the resulting small RNAs guide degradation or repression of matching transcripts rather than broadly changing all gene expression. This lets a screen connect a selected gene’s reduced expression with a resulting biological phenotype.
The phenotype selected for scoring determines which biological consequences become visible. Changes in development, movement, fertility, or stress responses can indicate that reduced expression affects distinct processes, but each readout links the gene to function only through the observed response. Examining several phenotype classes can broaden functional interpretation.
Short lifecycle and experimental tractability are important because they make results efficient to generate and follow up. In a large screen, these features support systematic testing of many selected genes and allow researchers to pursue genes whose knockdown produces informative phenotypes. This efficiency helps connect initial observations with pathway or disease-relevant interpretation.
Researchers select genes, provide double-stranded RNA through a delivery approach such as feeding bacteria engineered to produce it, and allow the RNA interference machinery to act. They then examine treated worms for changes in development, movement, fertility, or stress responses. Comparing these outcomes across targets reveals gene-phenotype associations.
Engineered bacteria provide a practical source of the double-stranded RNA used for treatment. Feeding worms this bacterial material connects gene selection with exposure in a format suitable for systematic experiments. The approach supports high-throughput screening because the same general delivery strategy can be applied across many targeted genes before phenotype assessment.
C. elegans RNAi screening is especially useful for functional genomics, where researchers need to assign roles to genes at scale. Results can also support pathway mapping and target identification by highlighting genes associated with observable biological changes. Because some phenotypes are disease-relevant, the screen can help prioritize mechanisms for follow-up investigation.