Synchronized lineage tracing of cell membranes and nuclei by dual recombinases and dual fluorescent

Xueying Yang; Shun He; Xufeng Li; Zhihou Guo; Haichang Wang; Zhuonan Zhang; Xin Song; Ke Jia; Lingjuan He; Bin Zhou

doi:10.1016/j.jgg.2024.07.006

Volume 51 Issue 12

Dec. 2024

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Article Navigation > Journal of Genetics and Genomics > 2024 > 51(12): 1474-1484

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Synchronized lineage tracing of cell membranes and nuclei by dual recombinases and dual fluorescent

doi: 10.1016/j.jgg.2024.07.006

a. School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, Zhejiang 310024, China;
b. School of Life Sciences, Westlake University, Hangzhou, Zhejiang 310030, China;
c. New Cornerstone Science Laboratory, State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China;
d. School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China

Funds:

This work was supported by the National Key Research & Development Program of China (2021YFA0805100, 2023YFA1800700, 2019YFA0110403, 2019YFA0802000), the National Science Foundation of China (82088101, 32370885, 92368103, 32370897), the Westlake Education Foundation, and the Benyuan Charity Fund, Research Funds of Hangzhou Institute for Advanced Study (2022ZZ01015 and B04006C01600515).

Received Date: 2024-05-03
Accepted Date: 2024-07-05
Rev Recd Date: 2024-07-03

Available Online: 2025-06-05

Publish Date: 2024-07-10

Abstract

Abstract

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.
- Lineage tracing,
- Dual recombinases system,
- Cre-loxP,
- Dre-rox,
- Cell membrane and nucleus labeling,
- ProTracer system

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References(43)

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