Microrna Knockdown
Microrna Knockdown - Related Videos
01:22
MicroRNAs
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2019
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
06:53
A Simple Alternative to Stereotactic Injection for Brain Specific Knockdown of miRNA
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Cited by 6 •
2015
MicroRNAs play crucial roles in the brain and are potential targets for modeling neuro-degeneration. However, perturbing miRNA levels is challenging due to the short length of miRNA and inaccessibility of the brain tissue. This video presents a method for antagomir design and brain specific delivery using a neuropeptide in mice.
09:19
In ovo Expression of MicroRNA in Ventral Chick Midbrain
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Cited by 5 •
2013
Ectopic expression is one technique to elucidate the microRNAs role in brain development. However, targeting specific areas using in ovo electroporation is challenging. Here, we show an efficient way to selectively electroporate ventral and dorsal midbrain regions.
07:19
Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
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Cited by 22 •
2011
MicroRNAs (miRNAs) are important regulators of gene expression and have been shown to play a role in numerous biological processes. To better understand miRNA-UTR interactions, we have created a genome-wide collection of 3 UTR luciferase reporters paired with a novel luciferase gene and assay reagent, the LightSwitch system.
09:17
Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits
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Cited by 7 •
2018
This protocol provides a workflow on how to combine artificial microRNA-mediated RNA interference with optogenetics to stimulate specifically presynaptic boutons with reduced expression of selective gene(s) within intact neuronal circuits.
FAQs
The intervention focuses on the mature microRNA rather than directly altering its messenger RNA targets. Antisense oligonucleotides, inhibitors, or microRNA sponges bind the mature molecule and reduce its ability to guide the RNA-induced silencing complex. This shifts regulation away from microRNA-mediated repression and allows affected target genes to become more highly expressed.
A mature microRNA normally guides the RNA-induced silencing complex toward complementary messenger RNAs, contributing to repression of their expression. When that microRNA is blocked, the complex is less able to impose this repression on the corresponding targets. The resulting increase in target-gene expression helps connect a particular microRNA to the pathways it regulates.
These reagent types provide different ways to interfere with the same regulatory interaction: each is introduced to bind the mature microRNA and prevent its normal activity. Their shared purpose is to reduce microRNA function, while the selected approach determines how the experimental system presents that blockade. This enables researchers to test whether observed effects depend on a specific microRNA.
Changes in expression after knockdown can identify genes that were previously suppressed by the microRNA and place those genes within broader regulatory pathways. In neuronal systems, this information can clarify how microRNAs influence differentiation, synaptic plasticity, neuroinflammation, or responses to injury. The approach therefore links a small RNA regulator with specific cellular processes.
Researchers first select the microRNA whose regulatory role they want to examine, then introduce an antisense oligonucleotide, inhibitor, or sponge designed to bind its mature form. They can evaluate whether affected target genes show restored expression and use those changes to investigate the associated molecular pathway. This workflow connects targeted inhibition with measurable gene-regulation outcomes.
It is useful when researchers need to determine whether a particular microRNA contributes to neuronal differentiation, synaptic plasticity, neuroinflammation, or cellular responses to injury. Reducing the microRNA’s activity reveals which target genes and pathways become less repressed. Those results can distinguish regulatory contributions that would remain unclear from microRNA expression measurements alone.
By restoring expression of genes affected by microRNA repression, the method can expose molecular pathways associated with neurological disorders. Researchers can use these pathway-level findings to identify microRNAs that may influence disease-related regulation and to evaluate whether their targets represent potential therapeutic opportunities. Its value lies in connecting regulatory mechanisms with disease-relevant molecular changes.