Rnai Genetic Screen
Rnai Genetic Screen - Related Videos
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.
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.
08:50
Genetic Screens
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2023
Genetic screens are critical tools for defining gene function and understanding gene interactions. Screens typically involve mutating genes and then assessing the affected organisms for phenotypes of interest. The process can be “forward”, where mutations are generated randomly to identify unknown genes responsible for the phenotypes, or it can be “reverse”, where specific genes are targeted for mutation to observe what phenotypes are produced.Here, JoVE reviews various types of genetic...
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.
08:20
RNAi Screening for Host Factors Involved in Vaccinia Virus Infection using Drosophila Cells
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Cited by 10 •
2010
Novel host factors involved in viral infection can be identified through cell-based genome-wide loss of function RNAi screening. A Drosophila cell culture model is particularly amenable to this approach due to the ease and efficiency of RNAi. Here we demonstrate this technique using vaccinia virus as an example.
FAQs
Sequence complementarity determines which messenger RNA is targeted. A double-stranded RNA or small interfering RNA guide directs the RNA-induced silencing complex, or RISC, toward a matching transcript. RISC-mediated degradation lowers the available messenger RNA, while translational suppression reduces protein production. This targeted reduction allows investigators to relate altered gene expression to a measured biological phenotype.
The approach examines many selected genes through systematic expression reduction rather than focusing on a single candidate. Researchers can then compare how each reduction changes a defined outcome, such as cell survival, cytokine production, or pathogen replication. Patterns across these measurements help identify genes that control the phenotype and organize them within broader defense or infection-related pathways.
Host-factor screens reduce expression of cellular genes to identify factors required for microbial entry, replication, or immune signaling. Complementary screens examine pathogen genes and their virulence functions instead. Considering both sides can connect host defense mechanisms with microbial requirements, helping researchers distinguish processes that support infection from those that contribute to resistance or immune responses.
A typical workflow selects a set of genes, introduces corresponding double-stranded RNA or small interfering RNA guides, and allows the RNA interference machinery to reduce matching transcripts or suppress their translation. Researchers then measure a chosen phenotype across the gene set, such as survival, cytokine production, or pathogen replication, and identify reductions associated with meaningful changes.
Cell survival, cytokine production, and pathogen replication provide complementary views of the response. A survival measurement can indicate whether a gene supports cellular persistence, whereas cytokine output reflects immune signaling. Changes in pathogen replication can reveal host factors or microbial functions linked to infection. Selecting the readout determines which aspect of the phenotype the screen emphasizes.
Findings can be used to map defense pathways, identify host factors involved in microbial entry or replication, and characterize pathogen virulence functions. They also help prioritize therapeutic targets by connecting gene activity with disease-related outcomes. In immunology, the results may clarify how gene perturbation changes cytokine production or other measured immune phenotypes.