通过胚胎干细胞中的同源重组高效生成无选择基因的大鼠基因敲除模型。
Efficient generation of selection-gene-free rat knockout models by homologous recombination in ES cells.
作者信息
Lan He, Li Shuping, Guo Zihang, Men Hongsheng, Wu Yuanyuan, Li Ning, Bryda Elizabeth C, Capecchi Mario R, Wu Sen
机构信息
State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
Veterinary Pathobiology, Rat Resource and Research Center, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
出版信息
FEBS Lett. 2016 Oct;590(19):3416-3424. doi: 10.1002/1873-3468.12388. Epub 2016 Sep 18.
Embryonic stem cell (ES cell)-based rat knockout technology, although successfully developed in 2010, has seen very limited usage to date due to low targeting efficiency and a lack of optimized procedures. In this study, we performed gene targeting in ES cells from the Sprague-Dawley (SD) and the Fischer 344 (F344) rat strains using an optimized procedure and the self-excising neomycin (neo)-positive selection cassette ACN to successfully generate Leptin and Trp53 knockout rats that did not carry the selection gene. These results demonstrate that our simplified targeting strategy using ACN provides an efficient approach to knock out many other rat genes.
基于胚胎干细胞（ES细胞）的大鼠基因敲除技术，尽管在2010年就已成功开发，但由于靶向效率低且缺乏优化程序，迄今为止其应用非常有限。在本研究中，我们使用优化程序和自我切除新霉素（neo）阳性选择盒ACN，对来自Sprague-Dawley（SD）和Fischer 344（F344）大鼠品系的ES细胞进行基因靶向，成功生成了不携带选择基因的瘦素和Trp53基因敲除大鼠。这些结果表明，我们使用ACN的简化靶向策略为敲除许多其他大鼠基因提供了一种有效的方法。
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