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Related Concept Videos

siRNA - Small Interfering RNAs02:30

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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
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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...
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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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piRNA - Piwi-interacting RNAs02:57

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PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...
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MicroRNA-based Regulation of Picornavirus Tropism
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Nuclear Argonaute:miRNA complexes recognize target sequences within chromatin and silence gene expression.

Cristina Hofman, Jiaxin Hu, Rut Bryl

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    RNA interference (RNAi) occurs in the cytoplasm and nucleus. Researchers used chimeric eCLIP to find that let-7 microRNAs target HMGA2 mRNA in both cellular locations, repressing its expression.

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    Area of Science:

    • Molecular Biology
    • Gene Regulation
    • RNA Biology

    Background:

    • RNA interference (RNAi) is crucial for gene silencing in mammalian cells, primarily occurring in the cytoplasm.
    • MicroRNAs (miRNAs) and RNAi factors are also found in the nucleus, but their nuclear functions in gene regulation are not fully understood.

    Purpose of the Study:

    • To investigate the nuclear roles of RNAi factors and miRNAs in gene expression regulation.
    • To identify nuclear miRNA:RNA interactions and their impact on target gene expression using a novel experimental approach.

    Main Methods:

    • Chimeric enhanced crosslinking immunoprecipitation (eCLIP) was employed to identify Argonaute 2 (AGO2)-miRNA complexes and their associated chromatin targets.
    • Quantitative analysis of miRNA binding sites and target mRNA levels in both cytoplasmic and nuclear cellular compartments.

    Main Results:

    • Chimeric eCLIP identified abundant nuclear miRNAs and their chromatin-associated RNA targets.
    • High mobility group AT-Hook A (HMGA2) was identified as a primary target of let-7 miRNA-mediated binding.
    • Let-7 miRNA repressed HMGA2 mRNA expression in both the cytoplasm and nucleus, with minimal impact on splicing or transcription.

    Conclusions:

    • RNAi-mediated gene silencing is not exclusively a cytoplasmic process; it can initiate in the nucleus.
    • Nuclear miRNA:RNA interactions contribute to the regulation of endogenous gene expression, influencing RNA levels in both nuclear and cytoplasmic compartments.
    • Chimeric eCLIP is a valuable tool for discovering functional miRNA:RNA interactions in vivo.