Genome-editing Technologies for Gene and Cell Therapy
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TLDR
The mechanisms of different genome-editing strategies are presented and each of the common nuclease-based platforms, including zinc finger nucleases, transcription activator-like effector nucleases (TALENs), meganucleases, and the CRISPR/Cas9 system are described.About:
This article is published in Molecular Therapy.The article was published on 2016-03-01 and is currently open access. It has received 515 citations till now. The article focuses on the topics: Transcription activator-like effector nuclease & Genome editing.read more
Citations
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Journal ArticleDOI
CRISPR-Cas9 Structures and Mechanisms.
Fuguo Jiang,Jennifer A. Doudna +1 more
TL;DR: This review aims to provide an in-depth mechanistic and structural understanding of Cas9-mediated RNA-guided DNA targeting and cleavage and provides a framework for rational engineering aimed at altering catalytic function, guide RNA specificity, and PAM requirements and reducing off-target activity for the development of Cas 9-based therapies against genetic diseases.
Journal ArticleDOI
The next generation of CRISPR-Cas technologies and applications.
TL;DR: The CRISPR–Cas toolkit has been expanding to include single-base editing enzymes, targeting RNA and fusing inactive Cas proteins to effectors that regulate various nuclear processes, and the new advances are considerably improving the authors' understanding of biological processes and are propelling CRISpr–Cas-based tools towards clinical use in gene and cell therapies.
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CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
Shengdar Q. Tsai,Nhu T. Nguyen,Jose Malagon-Lopez,Ved V Topkar,Martin J. Aryee,J. Keith Joung +5 more
TL;DR: Circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) is described, a highly sensitive, sequencing-efficient in vitro screening strategy that outperforms existing cell-based or biochemical approaches for identifying CRISPR–Cas9 genome-wide off-target mutations.
Journal ArticleDOI
Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells
Benjamin P. Kleinstiver,Shengdar Q. Tsai,Michelle S. Prew,Nhu T. Nguyen,Moira M. Welch,Jose Malagon Lopez,Zachary R. McCaw,Martin J. Aryee,J. Keith Joung +8 more
TL;DR: It is reported that four to six bases at the 3′ end of the short CRISPR RNA (crRNA) used to program Cpf1 nucleases are insensitive to single base mismatches, but that many of the other bases in this region of the crRNA are highly sensitive to single or double substitutions.
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Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts.
TL;DR: Techniques for membrane disruption-based intracellular delivery from 1911 until the present achieve rapid, direct, and universal delivery of almost any cargo molecule or material that can be dispersed in solution.
References
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Journal ArticleDOI
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
Martin Jinek,Krzysztof Chylinski,Krzysztof Chylinski,Ines Fonfara,Michael H. Hauer,Jennifer A. Doudna,Emmanuelle Charpentier +6 more
TL;DR: This 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
Le Cong,Le Cong,F. Ann Ran,F. Ann Ran,David M. Cox,David M. Cox,Shuailiang Lin,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +15 more
TL;DR: 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 as discussed by the authors.
Multiplex Genome Engineering Using CRISPR/Cas Systems
Le Cong,F. A. Ran,David Benjamin Turitz Cox,Shuailiang Lin,Robert P. J. Barretto,Naomi Habib,Patrick D. Hsu,Xuebing Wu,Wenyan Jiang,Luciano A. Marraffini,Feng Zhang +10 more
TL;DR: Two different type II CRISPR/Cas systems are engineered and it is demonstrated that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.
Journal ArticleDOI
RNA-Guided Human Genome Engineering via Cas9
Prashant Mali,Luhan Yang,Kevin M. Esvelt,John Aach,Marc Güell,James E. DiCarlo,Julie E. Norville,George M. Church,George M. Church +8 more
TL;DR: The type II bacterial CRISPR system is engineer to function with custom guide RNA (gRNA) in human cells to establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
Journal ArticleDOI
CRISPR provides acquired resistance against viruses in prokaryotes
Rodolphe Barrangou,Christophe Fremaux,Hélène Deveau,Melissa Richards,Patrick Boyaval,Sylvain Moineau,Dennis A. Romero,Philippe Horvath +7 more
TL;DR: It is found that, after viral challenge, bacteria integrated new spacers derived from phage genomic sequences, and CRISPR provided resistance against phages, and resistance specificity is determined by spacer-phage sequence similarity.