Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.
Department of Pharmacology, Shiga University of Medical Science, Shiga, Japan.
Sci Rep. 2023 Dec 18;13(1):22483. doi: 10.1038/s41598-023-50053-z.
The Cre-LoxP system has been commonly used for cell-specific genetic manipulation. However, many Cre strains exhibit excision activity in unexpected cell types or tissues. Therefore, it is important to identify the cell types in which recombination takes place. Fibroblasts are a cell type that is inadequately defined due to a lack of specific markers to detect the entire cell lineage. Here, we investigated the Cre recombination induced by Col1α2-iCre, one of the most common fibroblast-mesenchymal Cre driver lines, by using a double-fluorescent Cre reporter line in which GFP is expressed when recombination occurs. Our results indicated that Col1α2-iCre activity was more extensive across cell types than previously reported: Col1α2-iCre-mediated recombination was found in not only cells of mesenchymal origin but also those of other lineages, including haematopoietic cells, myocardial cells, lung and intestinal epithelial cells, and neural cells. In addition, study of embryos revealed that recombination by Col1α2-iCre was observed in the early developmental stage before gastrulation in epiblasts, which would account for the recombination across various cell types in adult mice. These results offer more insights into the activity of Col1α2-iCre and suggest that experimental results obtained using Col1α2-iCre should be carefully interpreted.
Cre-LoxP 系统已被广泛用于细胞特异性基因操作。然而,许多 Cre 株在预期之外的细胞类型或组织中表现出切除活性。因此,确定发生重组的细胞类型非常重要。成纤维细胞是一种细胞类型,由于缺乏检测整个细胞谱系的特异性标记物,因此定义不充分。在这里,我们使用一种双荧光 Cre 报告系,当发生重组时 GFP 表达,研究了 Col1α2-iCre(最常见的成纤维细胞间充质 Cre 驱动系之一)诱导的 Cre 重组。我们的结果表明,Col1α2-iCre 的活性比以前报道的更广泛:Col1α2-iCre 介导的重组不仅发生在间充质来源的细胞中,也发生在其他谱系的细胞中,包括造血细胞、心肌细胞、肺和肠上皮细胞以及神经细胞。此外,胚胎研究表明,Col1α2-iCre 的重组发生在原肠胚形成前的早期胚胎外胚层发育阶段,这可以解释成年小鼠中各种细胞类型的重组。这些结果提供了对 Col1α2-iCre 活性的更深入了解,并提示使用 Col1α2-iCre 获得的实验结果应谨慎解释。