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Curbing Malaria: A New Hope through Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Technology

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      RNA-guided human genome engineering via Cas9.

      Bacteria and archaea have evolved adaptive immune defenses, termed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems, that use short RNA to direct degradation of foreign nucleic acids. Here, we engineer the type II bacterial CRISPR system to function with custom guide RNA (gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4% in induced pluripotent stem cells. We show that this process relies on CRISPR components; is sequence-specific; and, upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci. We also compute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons. Our results establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
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        RNA-guided genetic silencing systems in bacteria and archaea.

        Clustered regularly interspaced short palindromic repeat (CRISPR) are essential components of nucleic-acid-based adaptive immune systems that are widespread in bacteria and archaea. Similar to RNA interference (RNAi) pathways in eukaryotes, CRISPR-mediated immune systems rely on small RNAs for sequence-specific detection and silencing of foreign nucleic acids, including viruses and plasmids. However, the mechanism of RNA-based bacterial immunity is distinct from RNAi. Understanding how small RNAs are used to find and destroy foreign nucleic acids will provide new insights into the diverse mechanisms of RNA-controlled genetic silencing systems.
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          Cas9 as a versatile tool for engineering biology.

          RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein, will grant unprecedented control over cellular organization, regulation and behavior. Here we describe the Cas9 targeting methodology, detail current and prospective engineering advances and suggest potential applications ranging from basic science to the clinic.
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            Author and article information

            Affiliations
            [1 ]Animal Genetics Division, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India
            [2 ]Department of Community Medicine, Kalinga Institute of Medical Sciences, KIIT University, Bhubaneswar, India
            Author notes
            [* ] Corresponding author: Dr Ansuman Panigrahi, E-mail: dr.ansuman3@ 123456gmail.com
            Journal
            J Arthropod Borne Dis
            J Arthropod Borne Dis
            JAD
            JAD
            Journal of Arthropod-Borne Diseases
            Tehran University of Medical Sciences
            2322-1984
            2322-2271
            December 2017
            30 December 2017
            : 11
            : 4
            : 539-540
            5775160 jad-11-539
            Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences

            This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.

            Categories
            Letter to the Editor

            Infectious disease & Microbiology

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