Jardé Thierry, Kerr Genevieve, Akhtar Reyhan, Abud Helen E
Cancer Program, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.
The Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2018;1725:41-52. doi: 10.1007/978-1-4939-7568-6_4.
Mouse models of intestinal carcinogenesis are very powerful for studying the impact of specific mutations on tumour 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 visualised 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. This methodology provides a rapid platform for assessing the cellular changes induced by specific mutations in intestinal tissue.
肠道癌发生的小鼠模型对于研究特定突变对肿瘤起始和进展的影响非常有用。使用可随时间诱导的条件性等位基因，可以单独或组合研究突变。肠道癌发生的步骤很复杂，在细胞水平对活体动物进行成像具有挑战性。体外在三维类器官中培养肠道上皮组织的能力提供了一个易于操作的系统，该系统可以进行基因操作并易于可视化，以评估对活组织的特定生物学影响。在这里，我们描述了通过他莫昔芬诱导的Cre重组酶或短暂暴露于TAT-Cre蛋白来对转基因小鼠类器官中的基因进行条件性突变的方法。该方法为评估肠道组织中特定突变诱导的细胞变化提供了一个快速平台。
