Rnai Gene Knockdown
Rnai Gene Knockdown - Related Videos
06:57
RNAi Mediated Gene Knockdown and Transgenesis by Microinjection in the Necromenic Nematode Pristionchus pacificus
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Cited by 28 •
2011
In model organisms, transgenesis can manipulate gene functions while RNAi can knockdown specific mRNA transcripts 1-2. This protocol aims to illustrate the techniques needed to introduce stably transmitted DNA and transient double stranded RNA into the necromenic nematode Pristionchus pacificus for studies in evolutionary, developmental, and behavioral biology.
08:47
RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes
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Cited by 19 •
2012
In this protocol we combine RNAi-mediated gene silencing with an in vivo diuresis assay to study the effects knockdown of genes of interest has on mosquito fluid excretion.
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.
10:57
RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)
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Cited by 19 •
2013
In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.
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.
FAQs
Within RISC, sequence complementarity determines which messenger RNA is recognized by the siRNA guide. Argonaute then provides the key effector activity: it can cleave the matched transcript or contribute to its repression. Lower messenger RNA availability reduces production of the corresponding protein, allowing gene-level perturbation to be linked with observed biological changes.
siRNA-mediated knockdown emphasizes a designed small interfering RNA with complementarity to a selected transcript, whereas related microRNA pathways can repress translation and promote messenger RNA destabilization. This distinction matters when interpreting the molecular outcome: reduced protein production may reflect transcript cleavage, transcript destabilization, translational repression, or a combination of these effects.
Knockdown strength and specificity are central limitations. Incomplete silencing can leave enough target expression to produce a weak or ambiguous phenotype, while off-target silencing can introduce effects unrelated to the selected gene. Careful experimental controls are therefore needed to judge whether an observed result reflects the intended perturbation or an off-target effect.
In biology, researchers use the resulting change in phenotype to connect a selected gene with a biological pathway or process. If reducing expression alters a measurable phenotype, that observation can support a functional role for the gene, while interpretation still depends on accounting for incomplete or off-target silencing. This makes the approach useful for pathway analysis.
The approach is used in both cultured cells and whole organisms, giving researchers ways to examine gene-associated effects in different biological settings. Work in these systems can reveal changes relevant to biological pathways and can help model disease mechanisms. Results from either setting still depend on the completeness and specificity of silencing.
RNAi gene knockdown supports disease-mechanism studies and drug-target validation. Researchers can reduce expression of a selected gene, examine the resulting phenotypic effects, and ask whether that gene is associated with the process or target under investigation. The resulting evidence can connect molecular changes with biological or disease-related phenotypes while supporting evaluation of a target's relevance.