Transposon-mediated transgenesis, transgenic rescue, and tissue-specific gene expression in rodents and rabbits
Transposon-mediated transgenesis, transgenic rescue, and tissue-specific gene expression in rodents and rabbits
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DOI：
10.1096/fj.12-205526
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发表时间：
2013-03-01
期刊：
影响因子：
4.8
通讯作者：
Izsvak, Zsuzsanna
中科院分区：
文献类型：
--
作者：
Katter, Katharina;Geurts, Aron M.;Izsvak, Zsuzsanna
生殖系转基因是生物学途径功能研究和动物生物技术的重要步骤。我们已经建立了一个简单的，非病毒的协议，在三个重要的生物医学模式生物经常用于生理学研究。该方案基于超活性睡美人转座子系统SB 100 X，其在小鼠、大鼠和兔中分别以50-64、14-72和15%的频率可重复地促进转基因创始者的产生。SB 100 X介导的转基因整合与经典的原核注射或慢病毒介导的转基因相比，不太容易发生遗传镶嵌和基因沉默。该方法已成功应用于多种转基因和动物模型，并可用于产生具有单拷贝整合的创始人。转座子载体还允许产生具有由选择的启动子元件指定的组织特异性表达模式的转基因系，例如用于跟踪多巴胺能神经元的大鼠报告菌株。作为原理的证明，我们通过SB 100 X转基因挽救了驼色头巾高血压大鼠的先天性遗传缺陷。SB 100 X和piggyBac方案的并排比较显示，这两个系统是互补的，为基因组操作提供了新的机会。Katter，K.，Geurts，A. M.，霍夫曼，O.，马茨湖，兰达，五，希里皮湖莫雷诺角，Lazar，J.，巴希尔，S.，Zidek，V.，波波娃，E.，Jerchow，B.，Becker，K.，Devaraj，A.，沃尔特岛Grzybowksi，M.，Corbett，M.，Rangel Filho，A.，Hodges，M. R.，Bader，M.，伊维奇，Z.，Jacob，H. J.，Pravenec，M.，Bosze，Z.，Rulicke，T.，伊兹瓦克角转座子介导的转基因、转基因拯救和啮齿类动物和兔的组织特异性基因表达。FASEB J.27，930-941（2013）。www.fasebj.org
Germline transgenesis is an important procedure for functional investigation of biological pathways, as well as for animal biotechnology. We have established a simple, nonviral protocol in three important biomedical model organisms frequently used in physiological studies. The protocol is based on the hyperactive Sleeping Beauty transposon system, SB100X, which reproducibly promoted generation of transgenic founders at frequencies of 50-64, 14-72, and 15% in mice, rats, and rabbits, respectively. The SB100X-mediated transgene integrations are less prone to genetic mosaicism and gene silencing as compared to either the classical pronuclear injection or to lentivirus-mediated transgenesis. The method was successfully applied to a variety of transgenes and animal models, and can be used to generate founders with single-copy integrations. The transposon vector also allows the generation of transgenic lines with tissue-specific expression patterns specified by promoter elements of choice, exemplified by a rat reporter strain useful for tracking serotonergic neurons. As a proof of principle, we rescued an inborn genetic defect in the fawn-hooded hypertensive rat by SB100X transgenesis. A side-by-side comparison of the SB100X- and piggyBac-based protocols revealed that the two systems are complementary, offering new opportunities in genome manipulation.-Katter, K., Geurts, A. M., Hoffmann, O., Mates, L., Landa, V., Hiripi, L., Moreno, C., Lazar, J., Bashir, S., Zidek, V., Popova, E., Jerchow, B., Becker, K., Devaraj, A., Walter, I., Grzybowksi, M., Corbett, M., Rangel Filho, A., Hodges, M. R., Bader, M., Ivics, Z., Jacob, H. J., Pravenec, M., Bosze, Z., Rulicke, T., Izsvak, Z. Transposon-mediated transgenesis, transgenic rescue, and tissue-specific gene expression in rodents and rabbits. FASEB J. 27, 930-941 (2013). www.fasebj.org