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    Rna Interference Final

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    Parvathi Babu
    - RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression or translation. [Fire and Mello] discovered RNAi through gene silencing by double-stranded RNA in C. elegans. [Fire and Mello] were awarded the 2006 Nobel Prize in Physiology or Medicine for this discovery. RNAi has significance for understanding viral defense mechanisms, genome stability, development, epigenetics, and potential therapeutic applications.

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    Rna Interference Final

    Uploaded by

    Parvathi Babu
    38 views29 pages
    - RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression or translation. [Fire and Mello] discovered RNAi through gene silencing by double-stranded RNA in C. elegans. [Fire and Mello] were awarded the 2006 Nobel Prize in Physiology or Medicine for this discovery. RNAi has significance for understanding viral defense mechanisms, genome stability, development, epigenetics, and potential therapeutic applications.

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    - RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression or translation. [Fire and Mello] discovered RNAi through gene silencing by double-stranded RNA in C. elegans. [Fire and Mello] were awarded the 2006 Nobel Prize in Physiology or Medicine for this discovery. RNAi has significance for understanding viral defense mechanisms, genome stability, development, epigenetics, and potential therapeutic applications.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    38 views29 pages

    Rna Interference Final

    Uploaded by

    Parvathi Babu
    - RNA interference (RNAi) is a biological process in which RNA molecules inhibit gene expression or translation. [Fire and Mello] discovered RNAi through gene silencing by double-stranded RNA in C. elegans. [Fire and Mello] were awarded the 2006 Nobel Prize in Physiology or Medicine for this discovery. RNAi has significance for understanding viral defense mechanisms, genome stability, development, epigenetics, and potential therapeutic applications.

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    of 29

    RNAi

    Regulation of
    X Biological
    Function
    RNAi
    The Nobel Prize in Physiology or Medicine 2006 was awarded
    jointly to Andrew Z. Fire and Craig C. Mello "for their discovery of
    RNA interference - gene silencing by double-stranded RNA"
    Watson & Crick
    Nobel prize 1962
    Fire & Mello
    Roger Kornberg Nobel prize 2006
    Nobel prize 2006

    transcription translation
    DNA RNA protein
    replication
    The genome operates by copying instructions for the
    manufacture of proteins from DNA into RNA in the
    Arthur Kornberg nucleus of the cell, then sending the RNA messages
    to the protein synthesizing machinery in the
    Nobel prize 1959
    cytoplasm. RNA interference blocks the translation
    of messenger RNAs for a specific gene, leading to
    silencing of expression of that gene.
    miRNA Biogenesis
    pre-miRNA

    pri-miRNA
    miRNA

    The pre-miRNA molecule is then actively transported out of the nucleus into the
    cytoplasm by Exportin 5,
    a carrier protein.

    The Dicer enzyme then cuts 20-25 nucleotides from the base of the hairpin to
    release
    the mature miRNA.

    In plants, which lack Drosha homologues, pri- and pre-miRNA processing by Dicer
    probably takes
    place in the nucleus, and mature miRNA duplexes are exported to the cytosol by
    Exportin 5.
    Fire and Mello could present a series of straightforward conclusions in their
    study. The main results can be summed up as follows:

    ▪ silencing was triggered efficiently by injected dsRNA, but weakly or not at all by
    sense or antisense single-stranded RNAs.

    ▪ silencing was specific for an mRNA homologous to the dsRNA; other mRNAs
    were unaffected.

    ▪ the dsRNA had to correspond to the mature mRNA sequence; neither intron
    nor promoter sequences triggered a response. This indicated a
    posttranscriptional, presumably cytoplasmic mechanism.

    ▪ the targeted mRNA disappeared suggesting that it was degraded.

    ▪ only a few dsRNA molecules per cell were sufficient to accomplish full silencing.
    This indicated that the dsRNA was amplified and/or acted catalytically rather
    than stoichiometrically.

    ▪ the dsRNA effect could spread between tissues and even to the progeny,
    suggesting a transmission of the effect between cells.
    Overview of RNAi

    • The dicer enzymes produce siRNA from double-stranded RNA and mature miRNA from precursor
    miRNA.
    • miRNA or siRNA is bound to an argonaute enzyme and an effector complex is formed, either
    a RISC (RNA-induced silencing complex) or RITS (RNA-induced transcriptional silencing) complex.
    • RITS affects the rate of transcription by histone and DNA methylation, whereas RISC
    degrades mRNA to prevent it from being translated.
    RNAi timeline
    Significance of the discovery of RNAi
    1. RNAi protects against viral infections
    2. RNAi secures genome stability by keeping mobile elements silent
    3. RNAi-like mechanisms repress protein synthesis and regulate the
    development of organisms
    4. RNAi-like mechanisms keep chromatin condensed and suppress
    transcription
    5. RNAi offers a new experimental tool to repress genes specifically
    6. RNAi might be a useful approach in future therapeutics
    Nomenclature of miRNA
    ▪ mir-123 , mir-456 (indicate order of naming)
    ▪ mir-123 (pre-miRNA), miR-123 (mature –miRNA)
    ▪ miRNA-123a, miRNA-123b (closely related)
    ▪ mir-194-1 and mir-194-2 (different gene locations) lead to an
    identical mature miR-194
    ▪ Species: hsa (human), oar (sheep), v (viral), d (drosophila)
    ▪ microRNAs originate from opposite arms of the same pre-miRNA,
    they are denoted with a -3p or -5p suffix.
    ▪ miRNA123, mirRNA123* (relative expression of opposite arm of
    hairpin)
    Similarities between miRNA and siRNA
    ▪ Mature miRNAs are structurally similar to siRNA
    ▪ Produced from dsRNA
    ▪ miRNA and siRNA share the same cellular
    machinery downstream of their initial processing
    ▪ The dsRNA portion of is bound and cleaved by
    Dicer to produce the mature miRNA molecule that
    can be integrated into the RISC complex
    Overall difference between miRNA and siRNA

    miRNA (micro RNA)


    ▪ 21-23 nt
    ▪ Produced from genes and endogenous dsRNA after maturity
    ▪ Is expressed from a much longer RNA coding gene as a primary transcript (prei-
    miRNA to pre-miRNA to mi-RNA)
    ▪ binds at the end of the mRNA (UTR)
    ▪ Acts as a signal to prevent translation

    siRNA (small interfering RNA)


    ▪ 20-25 nt
    ▪ Produced from exogenous dsRNA
    ▪ Binds to the coding region of mRNA
    ▪ Physically prevents translation
    RNAi can be induced by:
    RNAi is transitive:
    silencing can
    travel in a 3' to
    5' direction on
    a specific
    mRNA target
    Applications of RNAi

    • Gene Silencing
    • Gene Therapy
    • Selection of possible targets for Tumour
    therapy
    • Better understanding of viral infections
    THANK
    YOU

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