Journal ArticleDOI
Meganucleases and DNA double-strand break-induced recombination : Perspectives for gene therapy
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TLDR
Current systems based on redesigned endonucleases will be presented, with a special emphasis on the recent advances in homing endonuclease engineering, and the main issues that will need to be addressed in order to bring this promising technology to the patient.Abstract:
Meganucleases are sequence-specific endonucleases recognizing large (>12 bp) sequence sites and several laboratories have used these proteins to induce highly efficient gene targeting in mammalian cells. The recent development of artificial endonucleases with tailored specificities has opened the door for a wide range of new applications, including therapeutic ones: redesigned endonucleases cleaving chosen sequences could be used to in gene therapy to correct mutated genes or introduce transgenes in chosen loci. Such "targeted" approaches markedly differ from current gene therapy strategies based on the random insertion of a complementing virus-borne transgene. As a consequence, they should bypass the odds of random insertion. Artificial fusion proteins including Zinc-Finger binding domains have provided important proofs of concept, however the toxicity of these proteins is still an issue. Today custom-designed homing endonucleases, the natural meganucleases, could represent an efficient alternative. After a brief description of the origin of the technology, current systems based on redesigned endonucleases will be presented, with a special emphasis on the recent advances in homing endonuclease engineering. Finally, we will discuss the main issues that will need to be addressed in order to bring this promising technology to the patient.read more
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Journal ArticleDOI
Chromosomal context and epigenetic mechanisms control the efficacy of genome editing by rare-cutting designer endonucleases
Fayza Daboussi,Mikhail Zaslavskiy,Laurent Poirot,Mariana Loperfido,Agnès Gouble,Valérie Guyot,Sophie Leduc,Roman Galetto,Sylvestre Grizot,Danusia Oficjalska,Christophe Perez,Fabien Delacôte,Aurélie Dupuy,Isabelle Chion-Sotinel,Diane Le Clerre,Céline Lebuhotel,Olivier Danos,Frédéric Lemaire,Kahina Oussedik,Frédéric Cédrone,Jean-Charles Epinat,Julianne Smith,Rafael J. Yáñez-Muñoz,George Dickson,Linda Popplewell,Taeyoung Koo,Thierry VandenDriessche,Marinee K. Chuah,Aymeric Duclert,Philippe Duchateau,Frédéric Pâques +30 more
TL;DR: A comprehensive analysis on the efficacy of 37 endonucleases derived from the quintessential I-CreI meganuclease that were specifically designed to cleave 39 different genomic targets revealed that the efficiency of targeted mutagenesis at a given chromosome locus is predictive of that of homologous gene targeting.
Journal ArticleDOI
RNA-dependent DNA endonuclease Cas9 of the CRISPR system: Holy Grail of genome editing?
TL;DR: The potential of the Cas9 RNA-guided DNA endonuclease as a novel tool for genome surgery is highlighted, and possible constraints and future prospects are discussed.
Journal ArticleDOI
Limiting the Persistence of a Chromosome Break Diminishes Its Mutagenic Potential
Nicole Bennardo,Amanda Gunn,Amanda Gunn,Anita Cheng,Paul Hasty,Jeremy M. Stark,Jeremy M. Stark +6 more
TL;DR: Limiting the persistence of a DSB causes a reduction in the frequency of repair pathways that lead to significant genetic loss, and individual genetic factors play distinct roles during repair of non-cohesive DSB ends that are generated via co-expression of I-SceI with Trex2.
Journal ArticleDOI
Generation of redesigned homing endonucleases comprising DNA-binding domains derived from two different scaffolds
Sylvestre Grizot,Jean-Charles Epinat,Séverine Thomas,Aymeric Duclert,Sandra Rolland,Frédéric Pâques,Philippe Duchateau +6 more
TL;DR: The modularity of the chimeric DmoCre meganuclease is demonstrated, by successfully assembling mutants with locally altered specificities affecting both I-DmoI and I-CreI subdomains in order to create active meganucleases with altered Specificities.
Journal ArticleDOI
Homing endonucleases: from basics to therapeutic applications
TL;DR: The use of tailored HEs opens up new possibilities for gene therapy in patients with monogenic diseases that can be treated ex vivo, and several members of this enzyme family have been used as templates to engineer tools to cleave DNA sequences that differ from their original wild-type targets.
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