Targeted genome editing in pluripotent stem cells using zinc-finger nucleases.
Methods
; 53(4): 339-46, 2011 Apr.
Article
em En
| MEDLINE
| ID: mdl-21185378
ABSTRACT
Zinc-finger nucleases (ZFNs) are designer nucleases capable of cleaving a prespecified target DNA within complex genomes. ZFNs consist of a non-specific endonuclease domain fused to an engineered DNA-binding domain that tethers the nuclease activity to the chosen chromosomal site. The endonuclease-induced DNA double strand break triggers a cellular DNA damage response, resulting in double strand break repair by either accurate homologous recombination (HR) or error-prone non-homologous end-joining (NHEJ). Thus, ZFNs are powerful tools for targeted genome engineering in a variety of mammalian cell types, including embryonic (ESCs) and induced pluripotent stem cells (iPSCs). As a paradigm for genome editing in pluripotent stem cells, we describe the use of ZFNs in murine ESCs for generating knockout alleles by NHEJ without selection or by HR employing different selection schemes.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Dedos de Zinco
/
Genoma
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Desoxirribonucleases
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Células-Tronco Embrionárias
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Técnicas de Introdução de Genes
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Técnicas de Silenciamento de Genes
Limite:
Animals
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article