Sustained somatic gene inactivation by viral transfer of Cre recombinase
Sustained somatic gene inactivation by viral transfer of Cre recombinase
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DOI：
10.1038/nbt1196-1562
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发表时间：
1996-11-01
影响因子：
46.9
通讯作者：
Herz, J
中科院分区：
文献类型：
--
作者：
Rohlmann, A;Gotthardt, M;Herz, J
转基因和基因敲除小鼠已被证明是分析众多基因的生理相关功能的宝贵工具。然而，在某些情况下，不同组织、细胞类型或胚胎发育阶段对特定基因的不同需求所导致的多效性效应可能会由于复杂的表型或胚胎致死性而使分析复杂化。loxP/Cre介导的重组系统允许在小鼠中进行组织特异性基因靶向，可用于克服这些问题。目前方法的局限性是携带loxP标记基因的小鼠必须与表达Cre的转基因小鼠杂交。在适当的组织中重组酶以获得所需的基因重排。我们使用携带 Cre 重组酶的重组腺病毒来诱导肝脏中几乎定量的体细胞基因破坏。目标基因是多功能低密度脂蛋白受体相关蛋白 (LRP)，这是 α(2)-巨球蛋白和其他配体的细胞表面受体。腺病毒感染后 Cre 的瞬时表达产生了预测的基因重排，使肝脏中的 LRP 功能失活。重排发生在感染后 6 天内，并保持稳定至少 28 天，结果证明腺病毒 Cre 基因转移适合在小鼠中诱导长期、定量和时间控制的基因破坏。
Transgenic and knockout mice have proven invaluable tools for analyzing physiologically relevant functions of numerous genes. In some cases, however, pleiotropic effects that result from a variable requirement for a particular gene in different tissues, cell types, or stages of embryonic development may complicate the analysis due to a complex phenotype or embryonic lethality, The loxP/Cre-mediated recombination system, which allows tissue-specific gene targeting in the mouse, can be used to overcome these problems, A limitation of current methods is that a mouse carrying a loxP-tagged gene must be crossed with a transgenic mouse expressing the Cre recombinase in an appropriate tissue to obtain the desired gene rearrangement. We have used recombinant adenovirus carrying the Cre recombinase to induce virtually quantitative somatic cell gene disruption in the liver. The targeted gene was the multifunctional low-density lipoprotein receptor-related protein (LRP), a cell surface receptor for alpha(2)-macroglobulin and other ligands, Transient expression of Cre following adenoviral infection produced the predicted gene rearrangement, functionally inactivating LRP in the liver. Rearrangement occurred within 6 days after infection and remained stable for at least 28 days, the results demonstrate the suitability of adenoviral Cre gene transfer to induce long-term, quantitative, and temporally controlled gene disruption in the mouse.