Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.
Development and Stem Cells Program, Monash Biomedicine Discovery Institute, Clayton, VIC, Australia.
Methods Mol Biol. 2023;2691:55-69. doi: 10.1007/978-1-0716-3331-1_5.
Mouse models of intestinal carcinogenesis are very powerful tools for studying the impact of specific mutations on tumor initiation and progression. Mutations can be studied both singularly and in combination using conditional alleles that can be induced in a temporal manner. The steps in intestinal carcinogenesis are complex and can be challenging to image in live animals at a cellular level. The ability to culture intestinal epithelial tissue in three-dimensional organoids in vitro provides an accessible system that can be genetically manipulated and easily visualized to assess specific biological impacts in living tissue. Here, we describe methodology for conditional mutation of genes in organoids from genetically modified mice via induction of Cre recombinase induced by tamoxifen or by transient exposure to TAT-Cre protein and subsequent phenotyping of the organoids. This methodology provides a rapid platform for assessing the cellular changes induced by specific mutations in intestinal tissue.
肠道癌变的小鼠模型是研究特定突变对肿瘤起始和进展影响的非常有力的工具。可以使用条件等位基因来单独和组合研究突变，这些等位基因可以以时间方式诱导。肠道癌变的步骤很复杂，在活体动物中以细胞水平进行成像具有挑战性。在体外培养三维类器官中的肠上皮组织提供了一种易于操作的系统，可以对其进行遗传操作并轻松观察，以评估活体组织中的特定生物学影响。在这里，我们描述了通过他莫昔芬诱导 Cre 重组酶或通过短暂暴露于 TAT-Cre 蛋白来诱导，从而在来自基因修饰小鼠的类器官中条件性突变基因的方法，以及随后对类器官进行表型分析。这种方法为评估肠道组织中特定突变引起的细胞变化提供了快速平台。
