Electroporation-mediated Rnai
Electroporation-mediated Rnai - Related Videos
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.
07:56
Bacterial Delivery of RNAi Effectors: Transkingdom RNAi
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Cited by 18 •
2010
For development of RNA interference (RNAi)-based therapies, a novel strategy was developed, transkingdom RNAi (tkRNAi). This technology uses non-pathogenic bacteria to produce and deliver therapeutic short hairpin RNA (shRNA) into target cells. Here, tkRNAi was successfully applied for reversal of classical ABCB1-mediated multidrug resistance (MDR) of cancer cells.
10:12
Methods for Study of Neuronal Morphogenesis: Ex vivo RNAi Electroporation in Embryonic Murine Cerebral Cortex
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Cited by 6 •
2012
To conduct a rapid assessment of the function of genes in the development of cerebral cortex, we describe methods involving the ex vivo electroporation of plasmids co-expressing inhibitory RNA (RNAi) and GFP in murine embryonic cortex. This protocol is amenable to the study of various aspects of neurodevelopment such as neurogenesis, neuronal migration and neuronal morphogenesis including dendrite and axon outgrowth.
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.
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.
FAQs
The introduced small interfering RNA or double-stranded RNA directs RNA-induced silencing complexes toward messenger RNA with a complementary sequence. The complex then promotes degradation of that messenger RNA or blocks its translation, reducing production of the associated protein. This sequence-matching mechanism connects the delivered RNA to suppression of a selected gene rather than producing nonspecific inhibition.
Brief electrical pulses temporarily disrupt the cell membrane by creating pores through which small interfering RNA or double-stranded RNA can pass. Because the membrane disruption is temporary, the approach can deliver silencing molecules without requiring permanent genetic modification. This transient entry is especially useful when researchers need to alter gene activity during a defined developmental period.
The sequence of the introduced RNA determines the messenger RNA targeted by the silencing machinery. A molecule with complementarity to the selected transcript can guide suppression through messenger RNA degradation or translation blocking. Consequently, the experimental interpretation depends on linking the delivered sequence to the developmental gene under investigation, such as one involved in pattern formation or differentiation.
A typical experiment introduces sequence-specific small interfering RNA or double-stranded RNA into selected embryos, tissues, or cultured cells and then applies brief electrical pulses to promote entry across the membrane. The resulting gene suppression is examined in the treated region or population. This workflow supports rapid, localized perturbation while avoiding the need to generate a stable genetic modification.
Electroporation-mediated RNAi is useful when researchers need rapid suppression in a defined location rather than a permanently altered genetic state. Delivery can be directed to embryos, tissues, or cultured cells, allowing gene function to be examined during particular developmental events. This makes the method valuable for testing how reduced target-gene activity affects pattern formation, differentiation, or morphogenesis.
The method can reveal gene roles in pattern formation, cell differentiation, and morphogenesis by suppressing target transcripts in developing systems. Localized delivery helps associate a gene’s reduced activity with effects in a particular embryo or tissue, while the rapid nature of the perturbation supports functional analysis without stable genetic modification. These outcomes provide a way to connect gene activity with developmental form and organization.