Pronuclear injection-based mouse targeted transgenesis for reproducible and highly efficient transgene expression
Pronuclear injection-based mouse targeted transgenesis for reproducible and highly efficient transgene expression
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DOI：
10.1093/nar/gkq860
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发表时间：
2010-12-01
影响因子：
14.9
通讯作者：
Inoko, Hidetoshi
中科院分区：
文献类型：
--
作者：
Ohtsuka, Masato;Ogiwara, Sanae;Inoko, Hidetoshi
事实证明，小鼠转基因对于基因功能分析和人类疾病模型的产生是非常有价值的。我们在这里描述了一种基于原核注射的靶向转基因(PITT)系统的发展，涉及受精卵中的特定部位整合。将该系统应用于两个不同的基因组靶点，获得了包括荧光小鼠在内的一系列转基因品系，这些转基因品系可重复性地显示出强大、普遍和稳定的转基因表达。我们还证明，通过利用可重复性和高效的表达系统，PITT可以很容易地产生敲除小鼠。PITT系统绕过了传统随机整合转基因小鼠不可预测和不稳定的转基因表达问题，减少了产生转基因小鼠所需的时间、成本和精力，并可能适用于体内的功能获得和功能丧失研究。
Mouse transgenesis has proven invaluable for analysis of gene function and generation of human disease models. We describe here the development of a pronuclear injection-based targeted transgenesis (PITT) system, involving site-specific integration in fertilized eggs. The system was applied to two different genomic target loci to generate a series of transgenic lines including fluorescent mice, which reproducibly displayed strong, ubiquitous and stable transgene expression. We also demonstrated that knockdown mice could be readily generated by PITT by taking advantage of the reproducible and highly efficient expression system. The PITT system, which circumvents the problem of unpredictable and unstable transgene expression of conventional random-integration transgenic mice, reduces the time, cost and effort needed to generate transgenic mice, and is potentially applicable to both in vivo 'gain-of-function' and 'loss-of-function' studies.