Gene targeting methods for studying nuclear transport factors in mice
Gene targeting methods for studying nuclear transport factors in mice
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DOI：
10.1016/j.ymeth.2006.06.009
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发表时间：
2006-08-01
期刊：
影响因子：
4.8
通讯作者：
van Deursen, Jan M.
中科院分区：
文献类型：
--
作者：
Dawlaty, Meelad M.;van Deursen, Jan M.
基因工程小鼠已被广泛用于各种生命科学学科中的基因功能研究。然而，动物模型在核质运输领域的应用一直受到限制，主要是因为单个运输因子的干扰预计会严重放松对基本生物学过程的调控，导致胚胎在发育早期死亡。在老鼠体内，运输因子被敲除的早期研究已经证实了这一观点。最近的工作表明，亚型等位基因对于在生物体水平上研究基本基因非常有用。结合野生型和基因敲除等位基因，亚型等位基因可以用来产生一系列小鼠，在这些小鼠中，蛋白质的表达逐渐从正常减少到零。在这个系列中，通常有一个等位基因组合可以产生活着的小鼠，这些小鼠随着年龄的增长表现出明显的表型，可以用来研究蛋白质的生理相关性。在这篇文章中，我们提出了一种有效的方法来产生等位基因序列的小鼠。它涉及到使用一种多用途的基因靶向载体，这种载体可以产生一种亚型等位基因，这种等位基因也可以在小鼠体内转化为条件性和敲除等位基因。这种方法节省了时间，并在选择最合适的模型方面提供了灵活性，以便在生物体水平上研究核质机械的组件。(C)2006 Elsevier Inc.保留所有权利。
Genetically engineered mice have been widely used to study gene function in a variety of life-science disciplines. However, the use of animal models in the field of nucleocytoplasmic transport has been limited, mainly because disruption of individual transport factors is expected to deregulate basic biological processes so severely that the embryo dies at an early stage in development. Early studies in which transport factors were knocked out in mice have confirmed this notion. Recent work has shown that hypomorphic alleles are very useful for studying essential genes at the organismal level. In combination with wild-type and knockout alleles, hypomorphic alleles can be used to generate a series of mice in which the expression of a protein is gradually reduced from normal to zero. Within this series, there is often an allelic combination that yields liveborn mice that develop overt phenotypes as they age, and that can be used to study the physiological relevance of the protein. In this article, we present an efficient method for generating an allelic series of mice. It involves the use of a multipurpose gene-targeting vector that produces a hypomorphic allele that can also be converted into conditional and knockout alleles within the mouse. This method saves time and provides flexibility in terms of choosing the most appropriate model for studying components of the nucleocytoplasmic machinery at the organismal level. (c) 2006 Elsevier Inc. All rights reserved.