Genomic Rnai Library
Genomic Rnai Library - Related Videos
11:59
Competitive Genomic Screens of Barcoded Yeast Libraries
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Cited by 44 •
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We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.
20:59
Large Insert Environmental Genomic Library Production
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Cited by 18 •
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Construction of a fosmid library with environmental genomic DNA isolated from the vertical depth continuum of a seasonally hypoxic fjord is described. The resulting clone library is picked into 384-well plates and archived for downstream sequencing and functional screening by the application of an automated colony picking system.
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.
08:40
Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
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Cited by 1 •
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The Target ID Library is a plasmid-based, genome-wide collection of cloned cDNA used to identify miRNA targets. Here we demonstrate its use and application.
10:58
Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans
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Cited by 25 •
2012
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.
FAQs
Sequence complementarity determines which messenger RNA becomes the target of silencing. After the RNA interference reagent is processed, its guide strand directs the RNA-induced silencing complex to a matching transcript. This interaction can promote messenger RNA degradation or suppress translation, reducing expression of the selected gene and producing a phenotype that can be measured in the screen.
These reagent types differ in their molecular format, but all provide sequence information that can guide silencing toward a selected transcript. Double-stranded RNA and small interfering RNA are distinct reagent forms, while short hairpin RNA represents another design. Comparing results across these formats can help connect reduced expression of a gene with an observed biological phenotype.
The guide strand supplies the sequence-specific information used by the RNA-induced silencing complex to recognize a matching messenger RNA. Its complementarity links the molecular reagent to one target transcript rather than to an unrelated sequence. That targeting step is essential because the resulting degradation or translation suppression provides the connection between gene perturbation and phenotype.
A library extends RNA interference beyond isolated gene tests by organizing reagents that can reduce expression across many or all genes in an organism or cell system. Researchers can therefore examine numerous perturbations under a common screening strategy and compare their measurable phenotypes, helping identify genes that contribute to shared biological processes or responses.
A typical screen applies sequence-specific RNA interference reagents representing many target genes, allows the reagents to produce reduced expression, and measures the resulting biological phenotypes. Researchers then associate particular phenotypic changes with the corresponding gene perturbations. This high-throughput workflow converts a broad collection of molecular interventions into evidence about gene function.
These screens can identify genes involved in development, signaling, disease pathways, and cellular responses. The key output is a relationship between reducing expression of a particular gene and observing a measurable change in the biological system. Such relationships help researchers prioritize genes for further study within a pathway or broader cellular process.