Generation of redesigned homing endonucleases comprising DNA-binding domains derived from two different scaffolds.
Nucleic Acids Res
; 38(6): 2006-18, 2010 Apr.
Article
em En
| MEDLINE
| ID: mdl-20026587
ABSTRACT
Homing endonucleases have become valuable tools for genome engineering. Their sequence recognition repertoires can be expanded by modifying their specificities or by creating chimeric proteins through domain swapping between two subdomains of different homing endonucleases. Here, we show that these two approaches can be combined to create engineered meganucleases with new specificities. We demonstrate the modularity of the chimeric DmoCre meganuclease previously described, 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. Moreover these new engineered DmoCre variants appear highly specific and present a low toxicity level, similar to I-SceI, and can induce efficient homologous recombination events in mammalian cells. The DmoCre based meganucleases can therefore offer new possibilities for various genome engineering applications.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Desoxirribonucleases de Sítio Específico do Tipo I
/
Enzimas de Restrição do DNA
/
Proteínas de Ligação a DNA
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article