Reporter gene mouse lines are routinely used for studies related to functional genomics, proteomics, cell biology or cell-based drug screenings, and represent a crucial platform for in vivo research. In the generation of knock-in reporter lines, new gene targeting methods provide several advantages over the standard transgenic techniques. First of all, specific targeting of the genome allows expression of the reporter gene under controlled conditions, whether in a specific locus in the genome or in a “safe harbor” locus. Historically, the ROSA26 locus is used for gene knock-in strategies by homologous recombination in mouse embryonic stem cells. The other preferred place for integration of the reporter transgene in the mouse genome is the endogenous promoter of a target gene. In this study, we employed TALENs to generate a reporter fusion protein expressed from its native promoter. For monitoring DNA damage response, we generated a mouse line expressing a mCitrine-tagged version of the FANCD2 protein, involved in DNA damage response and repair, and the Fanconi anemia (FA) pathway. This model could be a valuable tool for in vivo investigation of DNA damage.

报告基因小鼠品系通常用于与功能基因组学，蛋白质组学，细胞生物学或基于细胞的药物筛选相关的研究，并且代表体内研究的重要平台。在敲入报告基因系的产生中，新的基因靶向方法提供了优于标准转基因技术的多种优势。首先，基因组的特异性靶向使报道基因在受控条件下表达，无论是在基因组中的特定位点还是在“安全港”位点。历史上，ROSA26基因座通过小鼠胚胎干细胞中的同源重组用于基因敲入策略。报道基因转基因在小鼠基因组中整合的另一个优选位置是靶基因的内源启动子。在这项研究中，我们利用TALENs生成了从其天然启动子表达的报告基因融合蛋白。为了监测DNA损伤反应，我们生成了表达mCitrine标签的FANCD2蛋白的小鼠品系，其参与DNA损伤反应和修复以及Fanconi贫血（FA）途径。该模型可能是体内研究DNA损伤的有价值的工具。