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CRISPR-Cas9 is an RNA-mediated adaptive immune system that protects bacteria and archaea from viruses or plasmids. Herein we discuss the recent development of CRISPR-Cas9 into a key technology for genome editing, targeting, and regulation in a wide range of organisms and cell types. It requires a custom designed single guide-RNA (sgRNA), a Cas9 endonuclease, and PAM sequences in the target region. The sgRNA-Cas9 complex binds to its target and creates a double-strand break (DSB) that can be repaired by non-homologous end joining (NHEJ) or by the homology-directed repair (HDR) pathway, modifying or permanently replacing the genomic target sequence. Additionally, we highlight recent advances in the repurposing of CRISPR-Cas9 for repression, activation, and loci imaging. In this review, we underline the current progress and the future potential of the CRISPR-Cas9 system towards biomedical, therapeutic, industrial, and biotechnological applications.
Author: | Yvonne Y Chen Affiliation: Department of Chemical and Biomolecular Engineering, University of California—Los Angeles, Los Angeles, CA 90095, USA |
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Edition/Format: | ![]() |
Publication: | Trends in Immunology, v36 n11 (201511): 667-669 |
Peer-reviewed | |
Database: | Copyright 2017 Elsevier B.V. All rights reserved |
Summary: |
Recent advances in T-cell therapy for cancer, viral infections, and autoimmune diseases highlight the broad therapeutic potential of T-cell engineering. However, site-specific genetic manipulation in primary human T cells remains challenging. Two recent studies describe efficient genome editing in T cells using CRISPR and TALEN approaches.
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Author: | A Stern Affiliation: Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.; L Keren; O Wurtzel; G Amitai; R Sorek |
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Edition/Format: | ![]() |
Publication: | Trends in genetics : TIG, 2010 Aug; 26(8): 335-40 |
Peer-reviewed | |
Database: | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. |
Other Databases: | ArticleFirst; WorldCat; British Library Serials |
Summary: |
The recently discovered prokaryotic immune system known as CRISPR (clustered regularly interspaced short palindromic repeats) is based on small RNAs ('spacers') that restrict phage and plasmid infection. It has been hypothesized that CRISPRs can also regulate self gene expression by utilizing spacers that target self genes. By analyzing CRISPRs from 330 organisms we found that one in every 250 spacers is self-targeting, and that such self-targeting occurs in 18% of all CRISPR-bearing organisms. However, complete lack of conservation across species, combined with abundance of degraded repeats near self-targeting spacers, suggests that self-targeting is a form of autoimmunity rather than a regulatory mechanism. We propose that accidental incorporation of self nucleic acids by CRISPR can incur an autoimmune fitness cost, and this could explain the abundance of degraded CRISPR systems across prokaryotes.
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