Yang Xueying, He Shun, Li Xufeng, Guo Zhihou, Wang Haichang, Zhang Zhuonan, Song Xin, Jia Ke, He Lingjuan, Zhou Bin
School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, Zhejiang 310024, China.
School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310030, China.
J Genet Genomics. 2024 Dec;51(12):1474-1484. doi: 10.1016/j.jgg.2024.07.006. Epub 2024 Jul 10.
Genetic lineage tracing has been widely employed to investigate cell lineages and fate. However, conventional reporting systems often label the entire cytoplasm, making it challenging to discern cell boundaries. Additionally, single Cre-loxP recombination systems have limitations in tracing specific cell populations. This study proposes three reporting systems utilizing Cre, Dre, and Dre+Cre mediated recombination. These systems incorporate tdTomato expression on the cell membrane and PhiYFP expression within the nucleus, allowing for clear observation of the nucleus and membrane. The efficacy of these systems is successfully demonstrated by labeling cardiomyocytes and hepatocytes. The potential for dynamic visualization of the cell membrane is showcased using intravital imaging microscopy or three-dimensional imaging. Furthermore, by combining this dual recombinase system with the ProTracer system, hepatocyte proliferation is traced with enhanced precision. This reporting system holds significant importance for advancing the understanding of cell fate studies in development, homeostasis, and diseases.
遗传谱系追踪已被广泛用于研究细胞谱系和命运。然而，传统的报告系统通常会标记整个细胞质，这使得辨别细胞边界具有挑战性。此外，单一的Cre-loxP重组系统在追踪特定细胞群体方面存在局限性。本研究提出了三种利用Cre、Dre和Dre+Cre介导的重组的报告系统。这些系统在细胞膜上整合了tdTomato表达，在细胞核内整合了PhiYFP表达，从而能够清晰地观察细胞核和细胞膜。通过标记心肌细胞和肝细胞成功证明了这些系统的有效性。使用活体成像显微镜或三维成像展示了细胞膜动态可视化的潜力。此外，通过将这种双重组酶系统与ProTracer系统相结合，能够更精确地追踪肝细胞增殖。该报告系统对于推进对发育、稳态和疾病中细胞命运研究的理解具有重要意义。
