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      CRISPR/Cas: From Tumor Gene Editing to T Cell-Based Immunotherapy of Cancer

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          Abstract

          The clustered regularly interspaced short palindromic repeats system has demonstrated considerable advantages over other nuclease-based genome editing tools due to its high accuracy, efficiency, and strong specificity. Given that cancer is caused by an excessive accumulation of mutations that lead to the activation of oncogenes and inactivation of tumor suppressor genes, the CRISPR/Cas9 system is a therapy of choice for tumor genome editing and treatment. In defining its superior use, we have reviewed the novel applications of the CRISPR genome editing tool in discovering, sorting, and prioritizing targets for subsequent interventions, and passing different hurdles of cancer treatment such as epigenetic alterations and drug resistance. Moreover, we have reviewed the breakthroughs precipitated by the CRISPR system in the field of cancer immunotherapy, such as identification of immune system-tumor interplay, production of universal Chimeric Antigen Receptor T cells, inhibition of immune checkpoint inhibitors, and Oncolytic Virotherapy. The existing challenges and limitations, as well as the prospects of CRISPR based systems, are also discussed.

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          Most cited references216

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          Hallmarks of Cancer: The Next Generation

          The hallmarks of cancer comprise six biological capabilities acquired during the multistep development of human tumors. The hallmarks constitute an organizing principle for rationalizing the complexities of neoplastic disease. They include sustaining proliferative signaling, evading growth suppressors, resisting cell death, enabling replicative immortality, inducing angiogenesis, and activating invasion and metastasis. Underlying these hallmarks are genome instability, which generates the genetic diversity that expedites their acquisition, and inflammation, which fosters multiple hallmark functions. Conceptual progress in the last decade has added two emerging hallmarks of potential generality to this list-reprogramming of energy metabolism and evading immune destruction. In addition to cancer cells, tumors exhibit another dimension of complexity: they contain a repertoire of recruited, ostensibly normal cells that contribute to the acquisition of hallmark traits by creating the "tumor microenvironment." Recognition of the widespread applicability of these concepts will increasingly affect the development of new means to treat human cancer. Copyright © 2011 Elsevier Inc. All rights reserved.
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            A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.

            Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
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              Multiplex genome engineering using CRISPR/Cas systems.

              Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.
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                Author and article information

                Contributors
                Journal
                Front Immunol
                Front Immunol
                Front. Immunol.
                Frontiers in Immunology
                Frontiers Media S.A.
                1664-3224
                29 September 2020
                2020
                : 11
                : 2062
                Affiliations
                [1] 1Student Research Committee, Shahid Beheshti University of Medical Sciences , Tehran, Iran
                [2] 2Department of Biotechnology, College of Science, University of Tehran , Tehran, Iran
                [3] 3Hematology Department, Faculty of Medical Sciences, Tarbiat Modares University , Tehran, Iran
                [4] 4Oncopathology Research Center, Iran University of Medical Sciences , Tehran, Iran
                [5] 5Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences , Tehran, Iran
                Author notes

                Edited by: Rayne Rouce, Baylor College of Medicine, United States

                Reviewed by: Hongbin Wang, California Northstate University, United States; Chiara Bonini, Vita-Salute San Raffaele University, Italy

                *Correspondence: Masoud Soleimani, soleim_m@ 123456modares.ac.ir

                These authors share first co-authorship

                ORCID: Mohammadreza Azangou-Khyavy, orcid.org/0000-0002-4543-4633; Mobina Ghasemi, orcid.org/0000-0003-0752-2410; Javad Khanali, orcid.org/0000-0002-9853-454X; Melika Boroomand-Saboor, orcid.org/0000-0001-9031-3454; Monire Jamalkhah, orcid.org/0000-0002-0737-6313; Masoud Soleimani, orcid.org/0000-0003-1972-7771; Jafar Kiani, orcid.org/0000-0002-5907-5846

                This article was submitted to Cancer Immunity and Immunotherapy, a section of the journal Frontiers in Immunology

                Article
                10.3389/fimmu.2020.02062
                7553049
                33117331
                59a38a48-ec14-42c9-ba8a-9d43cbf40a24
                Copyright © 2020 Azangou-Khyavy, Ghasemi, Khanali, Boroomand-Saboor, Jamalkhah, Soleimani and Kiani.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 05 March 2020
                : 29 July 2020
                Page count
                Figures: 4, Tables: 0, Equations: 0, References: 216, Pages: 19, Words: 0
                Categories
                Immunology
                Review

                Immunology
                crispr,cancer treatment,gene therapy,cancer immunotherapy,car t cell therapy,genome-wide screening assays,oncolytic virotherapy

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