Novel double promoter approach for identification of transgenic animals: A tool for in vivo analysis of gene function and development of gene-based therapies
Novel double promoter approach for identification of transgenic animals: A tool for in vivo analysis of gene function and development of gene-based therapies
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DOI：
10.1002/mrd.10137
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发表时间：
2002-08-01
影响因子：
2.5
通讯作者：
Shi, YB
中科院分区：
文献类型：
--
作者：
Fu, LZ;Buchholz, D;Shi, YB
脊椎动物遗传学的进步使得通过基因敲除和转基因分析来研究发育中的动物的基因功能成为可能。这些进步鼓励了基因疗法的发展，通过引入外源基因来增强和/或取代功能失调或缺失的基因。然而，在脊椎动物中，这种分析通常涉及冗长的转基因动物筛选，例如基于pcr的基因型测定。在这里，我们报道了在转基因非洲爪蟾蝌蚪中使用双启动子质粒携带感兴趣的转基因和结晶蛋白启动子驱动的绿色荧光蛋白（GFP）。这一策略允许一个简单的检查在眼睛中的绿色荧光蛋白的存在，以确定转基因动物。基于pcr的基因分型和功能表征证实，所有在眼睛中表达GFP的动物确实携带所需的启动子/转基因单位。因此，这种和其他类似载体的使用将极大地改进当前的转基因方案，减少鉴定转基因动物的时间和成本。(C) 2002 Wiley-Liss, Inc。
Advances in vertebrate genetics have allowed studies of gene function in developing animals through gene knockout and transgenic analyses. These advances have encouraged the development of gene-based therapies through introduction of exogenous genes to enhance and/or replace dysfunctional or missing genes. However, in vertebrates, such analyses often involve tedious screening for transgenic animals, such as PCR-based genotype determinations. Here, we report the use of double-promoter plasmids carrying the transgene of interest and the crystallin-promotor-driven Green fluorescent protein (GFP) in transgenic Xenopus laevis tadpoles. This strategy allows a simple examination for the presence of GFP in the eyes to identify transgenic animals. PCR-based genotyping and functional characterization confirms that all animals expressing GFP in the eyes indeed carry the desired promoter/transgene units. Thus, the use of this and other similar vectors should dramatically improve current transgenesis protocols and reduce the time and cost for identifying transgenic animals. (C) 2002 Wiley-Liss, Inc.