State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
School of Life Science and Technology, Shanghai Tech University, Shanghai 201210, China.
J Biol Chem. 2020 May 8;295(19):6413-6424. doi: 10.1074/jbc.REV120.011631. Epub 2020 Mar 25.
Site-specific recombinases, such as Cre, are a widely used tool for genetic lineage tracing in the fields of developmental biology, neural science, stem cell biology, and regenerative medicine. However, nonspecific cell labeling by some genetic Cre tools remains a technical limitation of this recombination system, which has resulted in data misinterpretation and led to many controversies in the scientific community. In the past decade, to enhance the specificity and precision of genetic targeting, researchers have used two or more orthogonal recombinases simultaneously for labeling cell lineages. Here, we review the history of cell-tracing strategies and then elaborate on the working principle and application of a recently developed dual genetic lineage-tracing approach for cell fate studies. We place an emphasis on discussing the technical strengths and caveats of different methods, with the goal to develop more specific and efficient tracing technologies for cell fate mapping. Our review also provides several examples for how to use different types of DNA recombinase-mediated lineage-tracing strategies to improve the resolution of the cell fate mapping in order to probe and explore cell fate-related biological phenomena in the life sciences.
位点特异性重组酶，如 Cre，是发育生物学、神经科学、干细胞生物学和再生医学等领域中用于遗传谱系追踪的一种广泛应用的工具。然而，一些遗传 Cre 工具的非特异性细胞标记仍然是该重组系统的技术限制，这导致了数据的误解，并在科学界引起了许多争议。在过去的十年中，为了提高遗传靶向的特异性和精度，研究人员同时使用两种或更多种正交重组酶来标记细胞谱系。在这里，我们回顾细胞追踪策略的历史，然后详细阐述最近开发的用于细胞命运研究的双遗传谱系追踪方法的工作原理和应用。我们重点讨论了不同方法的技术优势和注意事项，目的是开发更特异和高效的细胞命运映射追踪技术。我们的综述还提供了一些示例，说明了如何使用不同类型的 DNA 重组酶介导的谱系追踪策略来提高细胞命运映射的分辨率，以探测和探索生命科学中与细胞命运相关的生物学现象。
