Knockout mouse production assisted by Blm knockdown |
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Authors: | Mikiko FUKUDA Mayuko INOUE Daisuke MURAMATSU Hitoshi MIYACHI Yoichi SHINKAI |
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Institution: | 1)Cellular Memory Laboratory, RIKEN, Saitama 351-0198, Japan;2)CREST (Core Research for Evolutionary Science and Technology), Japan Agency for Medical Research and Development (AMED), Japan;3)Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan;4)Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan |
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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. |
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Keywords: | Blm Embryonic stem cells (ESCs) Gene targeting |
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