Knockout mouse production assisted by Blm knockdown.
J Reprod Dev
; 62(1): 121-5, 2016.
Article
em En
| MEDLINE
| ID: mdl-26598326
ABSTRACT
Production of knockout mice using targeted embryonic stem cells (ESCs) is a powerful approach for investigating the function of specific genes in vivo. Although the protocol for gene targeting via homologous recombination (HR) in ESCs is already well established, the targeting efficiency varies at different target loci and is sometimes too low. It is known that knockdown of the Bloom syndrome gene, BLM, enhances HR-mediated gene targeting efficiencies in various cell lines. However, it has not yet been investigated whether this approach in ESCs is applicable for successful knockout mouse production. Therefore, we attempted to answer this question. Consistent with previous reports, Blm knockdown enhanced gene targeting efficiencies for three gene loci that we examined by 2.3-4.1-fold. Furthermore, the targeted ESC clones generated good chimeras and were successful in germline transmission. These data suggest that Blm knockdown provides a general benefit for efficient ESC-based and HR-mediated knockout mouse production.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Células-Tronco Embrionárias
/
RecQ Helicases
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Técnicas de Silenciamento de Genes
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article