Production of Germline Transgenic Prairie Voles (Microtus ochrogaster) Using Lentiviral Vectors
Production of Germline Transgenic Prairie Voles (Microtus ochrogaster) Using Lentiviral Vectors
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DOI：
10.1095/biolreprod.109.077529
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发表时间：
2009-12-01
影响因子：
3.6
通讯作者：
Young, Larry J.
中科院分区：
文献类型：
--
作者：
Donaldson, Zoe R.;Yang, Shang-Hsun;Young, Larry J.
对替代模式生物的研究已经对更广泛使用的动物模型（如实验室大鼠和小鼠）所不显示的行为和生理特征的调节产生了巨大的见解。特别是，比较方法往往利用最适合研究特定现象的物种。例如，对社会性一夫一妻制的草原田鼠和一夫多妻制的草地田鼠的比较研究有助于理解社会联系的遗传和神经生物学基础。然而，对草原田鼠等不太常用生物的实验室研究因缺乏遗传工具（包括操纵基因组的能力）而受到限制。在这里，我们表明，慢病毒载体介导的转基因是一种快速和有效的方法，在替代实验室啮齿动物生殖系转基因。将表达绿色荧光蛋白（GFP）的慢病毒载体注射到23个单细胞胚胎的卵黄周隙中，产生了3个活的后代（13%），其中一个（33%）含有由人泛素-C启动子驱动的GFP转基因的种系整合。相比之下，移植23个未注射的胚胎产生了6个活的后代（26%）。绿色荧光蛋白存在于检查的所有组织中，并在脑中广泛表达。GFP转基因是可遗传的，并且至少在F（2）代之前稳定表达。这项技术有可能允许在草原田鼠的特定基因候选人的调查，并提供了一个通用的协议，追求在许多不同的啮齿动物物种的生殖系转基因操作。
The study of alternative model organisms has yielded tremendous insights into the regulation of behavioral and physiological traits not displayed by more widely used animal models, such as laboratory rats and mice. In particular, comparative approaches often exploit species ideally suited for investigating specific phenomenon. For instance, comparative studies of socially monogamous prairie voles and polygamous meadow voles have been instrumental toward gaining an understanding of the genetic and neurobiological basis of social bonding. However, laboratory studies of less commonly used organisms, such as prairie voles, have been limited by a lack of genetic tools, including the ability to manipulate the genome. Here, we show that lentiviral vector-mediated transgenesis is a rapid and efficient approach for creating germline transgenics in alternative laboratory rodents. Injection of a green fluorescent protein (GFP)-expressing lentiviral vector into the perivitelline space of 23 single-cell embryos yielded three live offspring (13%), one of which (33%) contained germline integration of a GFP transgene driven by the human ubiquitin-C promoter. In comparison, transfer of 23 uninjected embryos yielded six live offspring (26%). Green fluorescent protein is present in all tissues examined and is expressed widely in the brain. The GFP transgene is heritable and stably expressed until at least the F(2) generation. This technology has the potential to allow investigation of specific gene candidates in prairie voles and provides a general protocol to pursue germline transgenic manipulation in many different rodent species.