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
Citations
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Journal ArticleDOI
Genome Engineering With Zinc-Finger Nucleases
TL;DR: The history of ZFN development is reviewed, considerable progress has been made in methods for deriving zinc-finger sets for new genomic targets, but approaches to design and selection are still being perfected.
Journal ArticleDOI
Safe harbours for the integration of new DNA in the human genome
TL;DR: 'genomic safe harbours' are discussed — chromosomal locations where therapeutic transgenes can integrate and function in a predictable manner without perturbing endogenous gene activity and promoting cancer.
Patent
Tal effector-mediated dna modification
TL;DR: In this paper, the authors present a method for gene targeting with transcription activator-like effector nucleases (TALENS) and discuss its application in the field of bioinformatics.
Journal ArticleDOI
Plant Genome Engineering with Sequence-Specific Nucleases
TL;DR: Genome engineering promises to advance basic plant research by linking DNA sequences to biological function and will enable plants' biosynthetic capacity to be harnessed to produce the many agricultural products required by an expanding world population.
Journal ArticleDOI
Meganucleases and other tools for targeted genome engineering: perspectives and challenges for gene therapy.
George H. Silva,Laurent Poirot,Roman Galetto,Julianne Smith,Guillermo Montoya,Philippe Duchateau,Frédéric Paques +6 more
TL;DR: These alternative approaches based on non-viral vectorization and/or targeted insertion aimed at achieving safer gene transfer are reviewed, with a special emphasis on megan nucleases, a family of naturally occurring rare-cutting endonucleases, and speculate on their current and future potential.
References
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Journal ArticleDOI
Crystal structure of the intein homing endonuclease PI-SceI bound to its recognition sequence.
TL;DR: The first X-ray structures of an intein–DNA complex, that of the two-domain homing endonuclease PI-SceI bound to its 36-base pair DNA substrate, have been determined in the presence and absence of Ca2+.
Journal ArticleDOI
Crystal structure of the thermostable archaeal intron-encoded endonuclease I-DmoI
TL;DR: The overall fold of I-DmoI is similar to that of two other LAGLIDADG proteins for which the structures are known, I-CreI and the endonuclease domain of PI-SceI, but the loops connecting the beta-strands differ most in the loops connected to the respective DNA target site sizes and geometries.