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    Crisper Cas 9

    Uploaded by

    Yash Mittal

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    © All Rights Reserved
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    Crisper Cas 9

    Uploaded by

    Yash Mittal
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    Crisper Cas 9

    Uploaded by

    Yash Mittal

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    © All Rights Reserved
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    CRISPR-Cas9

     It is a novel Gene editing or gene regulatory technique.


     It was recognized by Emmanuelle Charpentier and Jennifer Dounda (for which they
    awarded Nobel prize in chemistry in 2020)
    CRISPR: Clustered regularly interspaced short palindromic repeats
    o CRISPR found in genomes of prokaryotic organism (in bacterial and archaea). It found
    in approximately in 50% of sequenced bacterial genomes & 90% of sequenced archaea.
    o These sequences are originally derived from DNA fragments of bacteriophages that had
    previously infected the prokaryote. Later they are used by prokaryotes to detect and
    destroy DNA from similar bacteriophages during subsequent infections to prokaryotes.
    So these sequences play role as an antiviral (antiphase) defense system for prokaryotes
    and provide an acquired immunity to prevent reinfection of a bacteria by specific
    bacteriophages.
    o CRISPR complements the system of restriction endonucleases & methylases. CRISPR
    uses RNA-based targeting to bring the Cas9 nuclease to foreign or complementary DNA.
    CAS9: CRISPR associated protein 9
    o An endonuclease enzyme
    o It uses CRISPR sequences as a guide (as guide RNA).
    o Then recognizes specific DNA strands that are complementary to CRISPR sequence &
    cleave them.
    CRISPR CAS9
    o Both together are used to edit genes within organisms.
    o Within bacteria this CRISPR-RNA-Cas9 form complex, then they degrade and
    inactivates the targeted DNA.
    o The CRISPR system has been also adapted for use in eukaryotic cells, including human
    cells, where it acts as RNA-directed site-specific nuclease like it is in bacteria.
    o It has wide varieties of application in medicine, in basic biological research, food and
    farming industries.
     In medicine, it was used to experimentally to treat a patient with a genetic disorder
    (eg sickle cell disease), to treat viral infection like HIV etc.
     In basic biological research, CRISPR system used for gene deletion, gene editing,
    gene visualization, and even modulation of gene transcription.
    Thus, CRISPR has added an exciting new, highly efficient, and very specific technology
    for the manipulation of DNA and genetic analysis of mammalian cells.
    o A variant of the CRISPR-Cas system, C2c2, has been shown to site-specifically cleave
    RNA. This exciting discovery paves the way for potential specific alteration of mRNA
    levels in human cells.

    By: Dr Mukesh Gohel, NHLMMC, Ahmedabad


    Reference: Harper’s illustrated biochemistry 31st edition PN 435
    ***End***

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