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    Gene Silencing
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    Gene Silencing Experiments

    • RNAi for functional genomics in plants [View Experiment]
    • Top 10 Tips For Gene Silencing & Delivery [View Experiment]
    • Gene Knockdown Functional Assays [View Experiment]
    • Optimizing electroporation parameters for effective gene silencing in Jurkat cells [View Experiment]
    • Successful RNA Interference Experiments in Difficult-to-Transfect Cell Lines: Experimental workflow for gene silencing with Thermo Scientific Dharmacon Accell siRNA and mRNA detection using Thermo Scientific Verso QRT-PCR [View Experiment]
    • Optimized Purification of siRNA Oligonucleotides Using the WAVE® Oligo System [View Experiment]
    • siRNA-mediated off-target gene silencing triggered by a 7 nt complementation [View Experiment]
    • Targefect-siRNA transfection kit for RNA interference studies! [View Experiment]
    • Using the Experion™ Automated Electrophoresis System to Assess RNA Quality and Quantity in siRNA-Induced Gene Silencing Experiments [View Experiment]
    • Dissertation: Probabilistic Models for Gene Silencing Data [View Experiment]
    • Thesis: Investigating the Roles of Micrornas In Biotic Stress Responses And Functional Characterization Of A Novel Ztl-Type F-Box Protein Via Virus Induced Gene Silencing [View Experiment]
    Gene Silencing Background Information

    Definition

    Gene silencing is a general term describing epigenetic processes of gene regulation. The term gene silencing is generally used to describe the "switching off" of a gene by a mechanism other than genetic modification. That is, a gene which would be expressed (turned on) under normal circumstances is switched off by machinery in the cell.

    Basics

    Genes are regulated at either the transcriptional or post-transcriptional level.

    Transcriptional gene silencing is the result of histone modifications, creating an environment of heterochromatin around a gene that makes it inaccessible to transcriptional machinery (RNA polymerase, transcription factors, etc.).

    Post-transcriptional gene silencing is the result of mRNA of a particular gene being destroyed or blocked. The destruction of the mRNA prevents translation to form an active gene product (in most cases, a protein). A common mechanism of post-transcriptional gene silencing is RNAi.

    Both transcriptional and post-transcriptional gene silencing are used to regulate endogenous genes. Mechanisms of gene silencing also protect the organism's genome from transposons and viruses. Gene silencing thus may be part of an ancient immune system protecting from such infectious DNA elements.

    Genes may be silenced by DNA methylation during meiosis, as in the filamentous fungus Neurospora crassa.

    Topics of Interest

    RNA interference (RNAi) is a system within living cells that helps to control which genes are active and how active they are. Two types of small RNA molecules – microRNA (miRNA) and small interfering RNA (siRNA) – are central to RNA interference. RNAs are the direct products of genes, and these small RNAs can bind to specific other RNAs and either increase or decrease their activity, for example by preventing a messenger RNA from producing a protein. RNA interference has an important role in defending cells against parasitic genes – viruses and transposons – but also in directing development as well as gene expression in general.

    RNA silencing (also called as post-transcriptional gene silencing PTGS) refers to a family of gene silencing effects by which the expression of one or more genes is downregulated or entirely suppressed by the introduction of an antisense RNA molecule. The most common and well-studied example is RNA interference, in which endogenously expressed microRNA or exogenously derived small interfering RNA induces the degradation of complementary messenger RNA. It also plays an important part in defending plants against viruses. Enzymes detect double stranded RNA (that is not normally found in cells) and digest it into small pieces that are not able to cause disease.

    Post transcriptional gene silencing (PTGS) is a mechanism for sequence-specific RNA degradation in plants. The process was described first in transgenic Petunia. The scientists’ goal was to produce petunia plants with improved flower colors. To achieve this goal, they introduced additional copies of a gene encoding a key enzyme for flower pigmentation into petunia plants. Surprisingly, many of the petunia plants carrying additional copies of this gene did not show the expected deep purple or deep red flowers but carried fully white or partially white flowers. When the scientists had a closer look they discovered that both types of genes, the endogenous and the newly introduced transgenes, had been turned off. Because of this observation the phenomenon was first named “co-suppression of gene expression” but the molecular mechanism remained unknown.

    Virus-Induced Gene Silencing (VIGS): A few years later plant virologists made a similar observation. In their research they aimed towards improvement of resistance of plants against plant viruses. At that time it was known that plants expressing virus-specific proteins show enhanced tolerance or even resistance against virus infection. However, they also made the surprising observation that plants carrying only short regions of viral RNA sequences not coding for any viral protein showed the same effect. They concluded that viral RNA produced by transgenes can also attack incoming viruses and stop them from multiplying and spreading throughout the plant. They did the reverse experiment and put short pieces of plant gene sequences into plant viruses. Indeed, after infection of plants with these modified viruses the expression of the targeted plant gene was suppressed. They called this phenomenon “virus-induced gene silencing” or simply “VIGS”.

    Source: Wikipedia (All text is available under the terms of the GNU Free Documentation License and Creative Commons Attribution-ShareAlike License.)

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