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    Type VI CRISPR-Cas System Overview

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    Juan Pablo Unfried Huertas
    The document discusses the Cas13a/C2C2 type VI CRISPR system. It claims that Cas13a engages in collateral cleavage of nearby non-targeted RNAs upon recognition of its RNA target. This collateral cleavage allows Cas13a to detect specific RNA presence in vivo or in vitro. It works by expressing Cas13a and a 28 nucleotide spacer CRNRA guide, with activity higher for guides in accessible regions. Cas13a can also process its own pre-crRNA for multiplexed guide delivery and its deactivated form maintains RNA binding for applications like transcript imaging.

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    Type VI CRISPR-Cas System Overview

    Uploaded by

    Juan Pablo Unfried Huertas
    12 views9 pages
    The document discusses the Cas13a/C2C2 type VI CRISPR system. It claims that Cas13a engages in collateral cleavage of nearby non-targeted RNAs upon recognition of its RNA target. This collateral cleavage allows Cas13a to detect specific RNA presence in vivo or in vitro. It works by expressing Cas13a and a 28 nucleotide spacer CRNRA guide, with activity higher for guides in accessible regions. Cas13a can also process its own pre-crRNA for multiplexed guide delivery and its deactivated form maintains RNA binding for applications like transcript imaging.

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    Overview of the recently described Crispr Cas13a for RNA targetting

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    Type VI CRISPR-Cas System overview

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    The document discusses the Cas13a/C2C2 type VI CRISPR system. It claims that Cas13a engages in collateral cleavage of nearby non-targeted RNAs upon recognition of its RNA target. This collateral cleavage allows Cas13a to detect specific RNA presence in vivo or in vitro. It works by expressing Cas13a and a 28 nucleotide spacer CRNRA guide, with activity higher for guides in accessible regions. Cas13a can also process its own pre-crRNA for multiplexed guide delivery and its deactivated form maintains RNA binding for applications like transcript imaging.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    Download as pptx, pdf, or txt
    12 views9 pages

    Type VI CRISPR-Cas System Overview

    Uploaded by

    Juan Pablo Unfried Huertas
    The document discusses the Cas13a/C2C2 type VI CRISPR system. It claims that Cas13a engages in collateral cleavage of nearby non-targeted RNAs upon recognition of its RNA target. This collateral cleavage allows Cas13a to detect specific RNA presence in vivo or in vitro. It works by expressing Cas13a and a 28 nucleotide spacer CRNRA guide, with activity higher for guides in accessible regions. Cas13a can also process its own pre-crRNA for multiplexed guide delivery and its deactivated form maintains RNA binding for applications like transcript imaging.

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    © All Rights Reserved
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    TYPE VI CRISPR-CAS SYSTEM

    RNA TARGETTING WITH CAS13A/C2C2

    JUAN PABLO UNFRIED


    JOURNAL CLUB 14TH NOV. 2017
    CAS13A SYSTEM
    CAS13A SYSTEM

    • WHAT THEY CLAIM

    • On recognition of its RNA target, activated Cas13a engages in “collateral” cleavage of


    nearby nontargeted RNAs (due to distance of cutting domains  Doudna)

    • This crRNA-programmed collateral-cleavage activity allows Cas13a to detect the presence


    of a specific RNA in vivo by triggering programmed cell death or in vitro by nonspecific
    degradation of labeled RNA.
    CAS13A SYSTEM

    • HOW DOES IT WORK


    CAS13A SYSTEM
    CAS13A SYSTEM

    • WHAT THEY CLAIM

    • LwaCas13a (from Leptotrichia wadei) better than Lsh.

    • Comparable levels of KD as RNAi with improved specificity.

    • dCas13a catalytically inactive maintains targeted RNA binding activity.


    CAS13A SYSTEM

    • HOW DOES IT WORK


    • Expression of Cas13a + 28 nt spacer (crRNA)
    • Guide lenghts down to 20nts still support cleavage, below that, binding may still happen
    without catalytic activity

    HEK293FT LNCRNAS
    CAS13A SYSTEM

    • HOW DOES IT WORK

    • LwaCas13a activity is higher


    for guides located in regions of
    accesibility.

    • It processes its own pre-


    crRNA  multiplexed
    delivery of guides.
    DCAS13A SYSTEM

    • WHAT THEY CLAIM


    • Works as a
    reprogrammable RNA
    binding protein.

    • Works as a transcript
    imaging platform.

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