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    North South University: CRISPR-Cas9

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    ishita hossain
    The document discusses the CRISPR-Cas9 system. It describes how CRISPR sequences in prokaryotes detect and destroy DNA from viruses during infections, providing antiviral defense. The Cas9 enzyme uses CRISPR sequences to recognize and cleave specific DNA strands. This CRISPR-Cas9 technology can edit genes. It consists of a Cas9 enzyme that acts as molecular scissors, and a guide RNA that binds to DNA and guides Cas9 to cut at the right point in the genome. The CRISPR-Cas9 system has been used for efficient genome editing in various species and applications such as research, medicine, and biotechnology.

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    North South University: CRISPR-Cas9

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    ishita hossain
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    The document discusses the CRISPR-Cas9 system. It describes how CRISPR sequences in prokaryotes detect and destroy DNA from viruses during infections, providing antiviral defense. The Cas9 enzyme uses CRISPR sequences to recognize and cleave specific DNA strands. This CRISPR-Cas9 technology can edit genes. It consists of a Cas9 enzyme that acts as molecular scissors, and a guide RNA that binds to DNA and guides Cas9 to cut at the right point in the genome. The CRISPR-Cas9 system has been used for efficient genome editing in various species and applications such as research, medicine, and biotechnology.

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    The document discusses the CRISPR-Cas9 system. It describes how CRISPR sequences in prokaryotes detect and destroy DNA from viruses during infections, providing antiviral defense. The Cas9 enzyme uses CRISPR sequences to recognize and cleave specific DNA strands. This CRISPR-Cas9 technology can edit genes. It consists of a Cas9 enzyme that acts as molecular scissors, and a guide RNA that binds to DNA and guides Cas9 to cut at the right point in the genome. The CRISPR-Cas9 system has been used for efficient genome editing in various species and applications such as research, medicine, and biotechnology.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    47 views5 pages

    North South University: CRISPR-Cas9

    Uploaded by

    ishita hossain
    The document discusses the CRISPR-Cas9 system. It describes how CRISPR sequences in prokaryotes detect and destroy DNA from viruses during infections, providing antiviral defense. The Cas9 enzyme uses CRISPR sequences to recognize and cleave specific DNA strands. This CRISPR-Cas9 technology can edit genes. It consists of a Cas9 enzyme that acts as molecular scissors, and a guide RNA that binds to DNA and guides Cas9 to cut at the right point in the genome. The CRISPR-Cas9 system has been used for efficient genome editing in various species and applications such as research, medicine, and biotechnology.

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    © All Rights Reserved
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    Download as pdf or txt
    of 5

    NORTH SOUTH UNIVERSITY

    Department of Biochemistry & Microbiology


    Course: Advanced Molecular Biology and
    Biochemistry

    Assignment-5
    CRISPR-Cas9

    Group Name- ZAPS

    Group members:

    Puja Banik, ID-1915410670

    Zahin Islam Rafa, ID-1915360070

    Md. Abir Hossain, ID-1825444670

    Saiful Islam Rony, ID- 1915489670

    Date: 7th September, 2019


    Background

    CRISPR (clustered regularly interspaced short palindromic repeats) is a family of DNA


    sequences found within the genomes of prokaryotic organisms such as bacteria and archaea.
    These sequences are derived from DNA fragments of viruses that have previously infected the
    prokaryote and are used to detect and destroy DNA from similar viruses during subsequent
    infections. Hence these sequences play a key role in the antiviral defense system of prokaryotes.

    Cas9 (CRISPR-associated protein 9) is an enzyme that uses CRISPR sequences as a guide to


    recognize and cleave specific strands of DNA that are complementary to the CRISPR sequence.
    Cas9 enzymes together with CRISPR sequences form the basis of a technology known as
    CRISPR-Cas9 that can be used to edit genes within organisms. This editing process has a wide
    variety of applications including basic biological research, development of biotechnology
    products, and treatment of diseases.

    The CRISPR-Cas system is a prokaryotic immune system that confers resistance to foreign
    genetic elements such as those present within plasmids and phages that provides a form of
    acquired immunity. RNA harboring the spacer sequence helps Cas (CRISPR-associated) proteins
    recognize and cut foreign pathogenic DNA. Other RNA-guided Cas proteins cut foreign RNA.
    CRISPR are found in approximately 50% of sequenced bacterial genomes and nearly 90% of
    sequenced archaea.

    Recently CRISPR-Cas9 system has become a potential genome editing tool that enables
    geneticists and medical researchers to edit parts of the genome by removing, adding or altering
    sections of the DNA sequence.

    How does CRISPR-Cas9 system work?

    The CRISPR-Cas9 system consists of two key molecules that introduce a change into the DNA.
    These are, 1) Cas9, An enzyme which acts as a pair of ‘molecular scissors’ that can cut the two
    strands of DNA at a specific location in the genome so that bits of DNA can then be added or
    removed; 2) A piece of RNA called guide RNA (gRNA). This consists of a small piece of pre-
    designed RNA sequence (about 20 bases long) located within a longer RNA scaffold. The
    scaffold part binds to DNA and the pre-designed sequence ‘guides’ Cas9 to the right part of the
    genome. This makes sure that the Cas9 enzyme cuts at the right point in the genome. The guide
    RNA is designed to find and bind to a specific sequence in the DNA. The guide RNA has RNA
    bases that are complementary to those of the target DNA sequence in the genome. This means
    that, at least in theory, the guide RNA will only bind to the target sequence and no other regions
    of the genome. The Cas9 follows the guide RNA to the same location in the DNA sequence and
    makes a cut across both strands of the DNA. At this stage the cell recognizes that the DNA is
    damaged and tries to repair it.
    Figure 1: Mechanism of CRISPR-Cas9 system.

    Specificity of Cas9 Nucleases

    Because genome editing leads to permanent modifications within the genome, the targeting
    specificity of Cas9 nucleases is of particular concern, especially for clinical applications and
    gene therapy. A combination of in vitro and in vivo assays has been typically used to
    characterize the specificity of ZFNs and TALENs (Gabriel et al., 2011), but systematic analysis
    has remained challenging due to difficulties in synthesizing large libraries of proteins with
    varying sequence specificity. However, Cas9 target recognition is dictated by the Watson-Crick
    base-pairing interactions of an RNA guide with its DNA target, enabling experimentally
    tractable and systematic evaluation of the effect of guide RNA-target DNA mismatches on Cas9
    activity.

    Genome Editing Using CRISPR-Cas9 in Eukaryotic Cells

    To date, the Streptococcus pyogenes Cas9 (SpCas9) has been used broadly to achieve efficient
    genome editing in a variety of species and cell types, including human cell lines, bacteria,
    zebrafish, yeast, mouse, fruit fly, roundworm, rat, common crops, pig, and monkey. SpCas9 is
    also dramatically expanding the catalog of genetically tractable model organisms, for example,
    by enabling the introduction of multiplex mutations in cynomolgus monkeys.

    Applications of Cas9 in Research, Medicine, and Biotechnology

    Cas9 can be used to facilitate a wide variety of targeted genome engineering applications. The
    wild-type Cas9 nuclease has enabled efficient and targeted genome modification in many species
    that have been intractable using traditional genetic manipulation techniques. The ease of
    retargeting Cas9 by simply designing a short RNA sequence also enables large-scale unbiased
    genome editing experiments to probe gene function or elucidate causal genetic variants.

    Figure 2: Mechanisms of Natural CRISPR in Adaptive Immunity of Microbial Systems.


    Conclusion

    The latest generation of Cas9-based genome engineering tools is also based on components from
    the microbial antiphage defense system. It is highly likely that the future solutions for efficient
    and precise gene modification will be found in as of yet unexplored corners of the rich biological
    diversity of nature.

    References

    1. Hsu P. Development and Applications of CRISPR-Cas9 for Genome Engineering.


    2014;157:1262–78.
    2. What is CRISPR-Cas9? | Facts | yourgenome.org [Internet]. [cited 2019 Sep 7]. Available
    from: https://www.yourgenome.org/facts/what-is-crispr-cas9

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