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RNA interference

life science Maturity 9-11 Next article →

RNA interference, or RNAi, is a biological mechanism used to suppress gene expression. This process involves RNA molecules that stop specific genes from working. It can happen through transcriptional repression, which stops DNA from being read. It can also happen through translational repression, which stops proteins from being built. RNAi is a naturally occurring pathway found in many eukaryotes, which are organisms with complex cells. It serves as a vital defense system against parasitic nucleotide sequences like viruses or transposons. It also plays a major role in the development of living organisms.

RNAi-simplified
RNAi-simplified

The mechanism begins when double-stranded RNA (dsRNA) enters the cell's cytoplasm. This dsRNA can be exogenous, meaning it comes from outside the cell like a virus. It can also be endogenous, meaning it originates from within the cell's own genome. An enzyme called Dicer initiates the process by cleaving these long dsRNA molecules. Dicer acts like molecular scissors to produce short fragments called small interfering RNAs, or siRNAs. These siRNAs are typically 21 to 23 nucleotides in length.

2ffl-by-domain
2ffl-by-domain

Once the siRNAs are created, they must be loaded into a functional complex. A group called the RISC-Loading Complex, or RLC, helps manage this step. The RLC includes proteins like Dicer-2 and R2D2. A factor called TAF11 helps assemble this complex by increasing the binding affinity to siRNA by 10-fold. The siRNA is then unwound into two single strands: the passenger strand and the guide strand. The protein Argonaute 2 (Ago2) cleaves the passenger strand so it can be degraded. The guide strand is then incorporated into the RNA-induced silencing complex, known as RISC.

Argonaute 1u04 1ytu composite
Argonaute 1u04 1ytu composite

The RISC uses the guide strand to find and silence specific targets. The guide strand pairs with a complementary sequence on a target mRNA molecule. Once the match is found, Ago2 acts as a catalytic component to induce cleavage of the mRNA. This prevents the mRNA from acting as a template for protein translation. This specific type of destruction is called post-transcriptional silencing. In some organisms, this response can even spread systemically throughout the body.

RNAi-simplified
RNAi-simplified

There is another important type of molecule involved in this pathway called microRNA, or miRNA. While siRNAs usually come from outside the cell, miRNAs are encoded within the genome. They start as long primary transcripts called pri-miRNA in the nucleus. A microprocessor complex, including the enzyme Drosha, processes these into a 70-nucleotide stem-loop structure called pre-miRNA. This pre-miRNA is then exported to the cytoplasm to be processed by Dicer.

Microrna secondary structure
Microrna secondary structure

Unlike siRNAs, miRNAs often bind to the 3′ untranslated region (3′UTR) of an mRNA with imperfect complementarity. Instead of cutting the mRNA, they often block ribosomes from translating it into a protein.

Historically, scientists did not realize these processes were all the same. They used different names like co-suppression, quelling, or post-transcriptional gene silencing. The true identity of RNAi was revealed through the work of Andrew Fire and Craig Mello. They studied the nematode worm *Caenorhabditis elegans* and published their findings in 1998. Their discovery was so significant that they shared the 2006 Nobel Prize in Physiology or Medicine.

Drosophila melanogaster - side (aka)
Drosophila melanogaster - side (aka)

Today, RNAi is a powerful tool in both research and medicine. Scientists use synthetic dsRNA to perform large-scale screens to identify which genes are necessary for processes like cell division. It is also being developed for use in food, medicine, and even insecticides.

ShRNA Lentivirus
ShRNA Lentivirus

In humans, the regulation of miRNA is critical for health. As of 2023, the miRBase archive listed 28,645 miRNA entries across 271 species. Dysregulation of these pathways is linked to serious conditions. For example, changes in miRNA are associated with various cancers and neuropsychiatric disorders like Alzheimer's disease and autism.

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🖼️ Images & Media (9)
File:ShRNA Lentivirus.svg
ShRNA Lentivirus.svg
File:2ffl-by-domain.png
2ffl-by-domain.png
File:Microrna secondary structure.png
Microrna secondary structure.png
File:Argonaute 1u04 1ytu composite.png
Argonaute 1u04 1ytu composite.png
File:RNAi-simplified.png
RNAi-simplified.png
File:Rnai diagram retrovirology.png
Rnai diagram retrovirology.png
File:Drosophila melanogaster - side (aka).jpg
Drosophila melanogaster - side (aka).jpg
Vertical Timeline of RNAi's Use in Medicine.pdf
File:Rnai phenotype petunia crop.png
Rnai phenotype petunia crop.png
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