Rnai Gene Editing
Rnai Gene Editing - Related Videos
08:41
Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis
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Cited by 5 •
2020
Here, we describe methods for efficient pupal and adult injections in Nasonia vitripennis as accessible alternatives to embryo microinjection, enabling functional analysis of genes of interest using either RNA-silencing via RNA interference (RNAi) or gene knockout via CRISPR/Cas9 genome editing.
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.
16:19
Localized RNAi and Ectopic Gene Expression in the Medicinal Leech
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Cited by 5 •
2008
In this video, we show a procedure for an accurate biolistic delivery of reagents into live tissue with a novel miniature gene gun. We are knocking down the expression of the axon guidance molecule Netrin in leech embryos by delivering molecules of dsRNA into the ventral body wall and ganglia of single segments.
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.
02:30
A CRISPR-Cas9 Technique for Gene Editing in T Cells
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2025
This video demonstrates an assay for performing gene editing in human T cells using the CRISPR-Cas9 technology. A mixture of primary CD4+ and CD8+ T cells is combined with a CRISPR-Cas9 ribonucleoprotein complex, targeting specific genes for knockout. Upon electroporation, the sgRNA guides Cas9 to the target DNA sequence, creating precise cuts. These cuts are then repaired by the cell's non-homologous end-joining mechanism, leading to gene knockout.
FAQs
After an siRNA or miRNA guides the RNA-induced silencing complex, sequence complementarity directs the complex toward matching messenger RNA. This recognition determines which transcript is affected. The targeted mRNA may then be degraded, or its translation may be blocked, reducing production from that gene without requiring a change to its DNA sequence.
The interaction between the guide RNA and complementary messenger RNA is central to the outcome. In some cases, targeting promotes mRNA degradation; in others, it prevents the message from being translated. These distinct outcomes both lower gene expression, but they provide different ways to interpret how RNAi has altered cellular regulation.
RNAi changes gene output at the messenger RNA level rather than modifying the underlying DNA sequence. Consequently, an observed phenotype reflects reduced expression of the targeted gene, not necessarily a permanent alteration of its genome. This distinction matters when researchers interpret results, compare experimental approaches, or design gene-regulation studies.
Researchers can reduce expression from a selected gene and then examine the resulting cellular phenotype. If the phenotype changes after the targeted transcript is silenced, the result can help link that gene to a cellular process or pathway. This functional-genomics strategy supports investigation of gene roles without directly changing the DNA sequence.
A study begins by selecting a gene whose function requires investigation, then using an siRNA or miRNA-based approach to guide RISC toward its complementary messenger RNA. Researchers evaluate the resulting reduction in gene expression and relate it to a cellular phenotype. This workflow helps test gene function and interpret pathway involvement.
RNAi can help investigate disease pathways by reducing expression of genes suspected to contribute to a biological process. It also supports drug-target validation, because silencing a candidate gene can provide functional evidence about its relevance. In addition, the approach contributes to studies of potential therapies while preserving the distinction between gene silencing and permanent genome modification.