Variegation and silencing in a lentiviral-based murine transgenic model
Variegation and silencing in a lentiviral-based murine transgenic model
复制标题
DOI：
10.1007/s11248-009-9318-4
复制
发表时间：
2010-06-01
影响因子：
3
通讯作者：
Leo, Oberdan
中科院分区：
文献类型：
--
作者：
Baup, Delphine;Fraga, Laurent;Leo, Oberdan
基于慢病毒的构建体代表了转基因动物的最新发展。使用的方便性，以及相同的主干载体可用于下调内源性基因表达和产生过表达感兴趣基因的转基因动物的事实，已经激发了人们对这项技术越来越大的兴趣。在这项研究中，我们使用慢病毒传递系统来产生表达改变水平（上调或下调）感兴趣的基因的转基因小鼠。尽管这种基于慢病毒的方法导致了高水平的转基因和种系传播，但在大多数第一代和第二代小鼠系中观察到转基因表达的广泛差异。特别是，尽管整合物分离到单拷贝表达的小鼠系中，转基因表达似乎是表观遗传调控机制的目标，经常导致在特定组织（如血液外周淋巴细胞）中高表达和低表达转基因细胞共存。因此，可能需要建立和分析大量的小鼠系，以选择稳定的转基因系，使用这种基于慢病毒的方法表达感兴趣的基因。
Lentiviral based constructs represent a recent development in the generation of transgenic animals. The ease of use, and the fact that the same backbone vectors can be used to down-modulate endogenous gene expression and to produce transgenic animals overexpressing a gene of interest, have fuelled growing interest in this technology. In this study, we have used a lentiviral delivery system to generate transgenic mice expressing altered levels (up or downregulated) of a gene of interest. Although this lentiviral-based approach led to high levels of transgenesis and germ line transmission, a wide variation in transgene expression was observed in most first and second generation mouse lines. In particular, despite the segregation of integrants into single-copy expressing mouse lines, transgene expression appeared to be the target of epigenetic regulatory mechanism, often causing the coexistence of high and low transgene expressing cells within a given tissue such as blood peripheral lymphocytes. The establishment and analysis of large number of mouse lines may therefore be required to select a stable transgenic line with pancellular expression of a gene of interest using this lentiviral-based approach.