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.Additiona...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-lox P 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 td Tomato expression on the cell membrane and Phi YFP 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 threedimensional imaging.Furthermore,by combining this dual recombinase system with the Pro Tracer 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.展开更多
Stem cell research has become a hot topic in biology,as the understanding of stem cell biology can provide new insights for both regenerative medicine and clinical treatment of diseases.Accurately deciphering the fate...Stem cell research has become a hot topic in biology,as the understanding of stem cell biology can provide new insights for both regenerative medicine and clinical treatment of diseases.Accurately deciphering the fate of stem cells is the basis for understanding the mechanism and function of stem cells during tissue repair and regeneration.Cre-loxP-mediated recombination has been widely applied in fate mapping of stem cells for many years.However,nonspecific labeling by conventional cell lineage tracing strategies has led to discrepancies or even controversies in multiple fields.Recently,dual recombinase-mediated lineage tracing strategies have been developed to improve both the resolution and precision of stem cell fate mapping.These new genetic strategies also expand the application of lineage tracing in studying cell origin and fate.Here,we review cell lineage tracing methods,especially dual genetic approaches,and then provide examples to describe how they are used to study stem cell fate plasticity and function in vivo.展开更多
基金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)。
文摘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-lox P 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 td Tomato expression on the cell membrane and Phi YFP 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 threedimensional imaging.Furthermore,by combining this dual recombinase system with the Pro Tracer 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.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB19000000 and XDA16010507)the National Key Research and Development Program of China(2019YFA0110403 and 2019YFA0802000)National Natural Science Foundation of China(31730112,31625019,91849202,and 82008810001)。
文摘Stem cell research has become a hot topic in biology,as the understanding of stem cell biology can provide new insights for both regenerative medicine and clinical treatment of diseases.Accurately deciphering the fate of stem cells is the basis for understanding the mechanism and function of stem cells during tissue repair and regeneration.Cre-loxP-mediated recombination has been widely applied in fate mapping of stem cells for many years.However,nonspecific labeling by conventional cell lineage tracing strategies has led to discrepancies or even controversies in multiple fields.Recently,dual recombinase-mediated lineage tracing strategies have been developed to improve both the resolution and precision of stem cell fate mapping.These new genetic strategies also expand the application of lineage tracing in studying cell origin and fate.Here,we review cell lineage tracing methods,especially dual genetic approaches,and then provide examples to describe how they are used to study stem cell fate plasticity and function in vivo.
