体细胞中转座子介导的诱变：转座子-基因组DNA连接点的鉴定
Transposon-mediated mutagenesis in somatic cells: identification of transposon-genomic DNA junctions.
作者信息
Largaespada David A, Collier Lara S
机构信息
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN.
出版信息
Methods Mol Biol. 2008;435:95-108. doi: 10.1007/978-1-59745-232-8_7.
Understanding the genetic basis for tumor formation is crucial for treating cancer. Forward genetic screens using insertional mutagenesis technologies have identified many important tumor suppressor genes and oncogenes in mouse models of human cancer. Traditionally, retroviruses have been used for this purpose, allowing the identification of genes that can cause various forms of leukemia or lymphoma with murine leukemia viruses or mammary cancer with mouse mammary tumor viruses. Recently, the Sleeping Beauty transposon system has emerged as a tool for cancer gene discovery in mouse models of human cancer. Transposons mobilized in the mouse soma can insertionally mutate cancer genes, and the transposon itself serves as a molecular "tag," which facilitates candidate cancer gene identification. We provide an overview of some general issues related to use of Sleeping Beauty for cancer genetic studies and present here the polymerase chain reaction-based method for cloning transposon-tagged sequences from tumors.
了解肿瘤形成的遗传基础对于癌症治疗至关重要。利用插入诱变技术进行的正向遗传学筛选已在人类癌症小鼠模型中鉴定出许多重要的肿瘤抑制基因和癌基因。传统上，逆转录病毒一直用于此目的，可利用鼠白血病病毒鉴定能引发各种形式白血病或淋巴瘤的基因，以及利用小鼠乳腺肿瘤病毒鉴定引发乳腺癌的基因。最近，睡眠美杜莎转座子系统已成为人类癌症小鼠模型中癌症基因发现的一种工具。在小鼠体细胞中移动的转座子可通过插入使癌症基因发生突变，而转座子本身充当分子“标签”，便于鉴定候选癌症基因。我们概述了与使用睡眠美杜莎进行癌症遗传研究相关的一些一般问题，并在此介绍基于聚合酶链反应从肿瘤中克隆转座子标记序列的方法。
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