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Protein Eng Des Sel. 2011 Jan;24(1-2):27-31. doi: 10.1093/protein/gzq083. Epub 2010 Nov 3.

The I-CreI meganuclease and its engineered derivatives: applications from cell modification to gene therapy.

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1
Cellectis Genome Surgery, 102 Avenue Gaston Roussel, 93 235 Romainville Cedex, France. arnould@cellectis.com

Abstract

Meganucleases (MNs) are highly specific enzymes that can induce homologous recombination in different types of cells, including mammalian cells. Consequently, these enzymes are used as scaffolds for the development of custom gene-targeting tools for gene therapy or cell-line development. Over the past 15 years, the high resolution X-ray structures of several MNs from the LAGLIDADG family have improved our understanding of their protein-DNA interaction and mechanism of DNA cleavage. By developing and utilizing high-throughput screening methods to test a large number of variant-target combinations, we have been able to re-engineer scores of I-CreI derivatives into custom enzymes that target a specific DNA sequence of interest. Such customized MNs, along with wild-type ones, have allowed for exploring a large range of biotechnological applications, including protein-expression cell-line development, genetically modified plants and animals and therapeutic applications such as targeted gene therapy as well as a novel class of antivirals.

PMID:
21047873
DOI:
10.1093/protein/gzq083
[Indexed for MEDLINE]
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