Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA.
PLoS One
; 8(10): e76478, 2013.
Article
em En
| MEDLINE
| ID: mdl-24130776
ABSTRACT
Zinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs. Here we describe a simple, non-integrating strategy for generating KO pigs using ZFN-encoding mRNA. The interleukin-2 receptor gamma (IL2RG) gene was knocked out in porcine fetal fibroblasts using ZFN-encoding mRNAs, and IL2RG KO pigs were subsequently generated using these KO cells through somatic cell nuclear transfer (SCNT). The resulting IL2RG KO pigs completely lacked a thymus and were deficient in T and NK cells, similar to human X-linked SCID patients. Our findings demonstrate that the combination of ZFN-encoding mRNAs and SCNT provides a simple robust method for producing KO pigs without genomic integration.
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Base de dados:
MEDLINE
Assunto principal:
Dedos de Zinco
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Desoxirribonucleases
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Subunidade gama Comum de Receptores de Interleucina
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Técnicas de Inativação de Genes
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Fibroblastos
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article