Placenta-Specific Gene Activation and Inactivation Using Integrase-Defective Lentiviral Vectors With the Cre/LoxP System
Placenta-Specific Gene Activation and Inactivation Using Integrase-Defective Lentiviral Vectors With the Cre/LoxP System
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DOI：
10.1002/dvg.20563
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发表时间：
2009-12-01
期刊：
影响因子：
1.5
通讯作者：
Ikawa, Masahito
中科院分区：
文献类型：
--
作者：
Morioka, Yuka;Isotani, Ayako;Ikawa, Masahito
在过去的几十年里，转基因和基因敲除研究已经推进了我们对胚胎发育的遗传控制的理解。然而，突变小鼠的表型的解释可能是复杂的，因为常用的敲除方法修改胎儿和胎盘基因组。为了解决这个问题，我们以前开发了一个胎盘特异性基因操作系统的慢病毒载体转导的胚胎在囊胚阶段。在本研究中，通过结合Cre/LoxP系统，我们成功地证明了胎盘特异性基因的激活和失活的EGFP报告小鼠和Ets 2 floxed小鼠，分别。使用整合酶缺陷型慢病毒（IDLV）载体的瞬时表达减少了Cre表达的毒性作用，并且解决了较低浓度的嵌合重组和较高浓度的Cre重组酶的毒性作用的困境。我们还表明，胎盘特异性Ets 2中断导致胚胎死亡，并再次证实了在胎盘形成过程中的关键作用Ets 2。这项技术有助于在妊娠期间胎盘发育中进行基因功能分析。由于IDLV载体可以类似于野生型载体有效地扩增多种细胞类型，因此我们的IDLV-Cre策略对于广泛的应用是潜在有用的。创世纪47：793-798，2009。(C)2009威利-利斯公司
Transgenic and knockout studies have advanced our understanding of the genetic control of embryonic development over the past decades. However, interpretation of the phenotype of mutant mice is potentially complicated, since the commonly used knockout approach modifies both the fetal and placental genome. To circumvent this problem, we previously developed a placenta-specific gene manipulation system by lentiviral vector transduction of embryos at the blastocyst stage. In the present study, by combination with the Cre/LoxP system, we successfully demonstrate placenta-specific gene activation and inactivation in EGFP reporter mice and Ets2 floxed mice, respectively. Transient expression using integrase-defective lentiviral (IDLV) vectors diminished the toxic effect of Cre expression and solved the dilemma of mosaic recombination with lower concentrations and toxic effects with higher concentrations of Cre recombinase. We also show that placenta-specific Ets2 disruption causes embryonic lethality and reconfirmed the critical role of Ets2 during placentation. This technology facilitates both gain and loss of gene function analyses in placental development during pregnancy. Since IDLV vectors can efficiently transduce a variety of cell types similarly to wild-type vectors, our IDLV-Cre strategy is potentially useful for a wide range of applications. genesis 47:793-798, 2009. (C) 2009 Wiley-Liss, Inc.