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Triple-cell lineage tracing by a dual reporter on a single allele.
Journal of Biological Chemistry ( IF 4.0 ) Pub Date : 2019-11-26 , DOI: 10.1074/jbc.ra119.011349
Kuo Liu 1, 2 , Muxue Tang 1 , Hengwei Jin 1 , Qiaozhen Liu 1 , Lingjuan He 1 , Huan Zhu 1 , Xiuxiu Liu 1 , Ximeng Han 1, 2 , Yan Li 1 , Libo Zhang 1 , Juan Tang 1 , Wenjuan Pu 1 , Zan Lv 1 , Haixiao Wang 1 , Hongbin Ji 1 , Bin Zhou 2, 3
Affiliation  

Genetic lineage tracing is widely used to study organ development and tissue regeneration. Multicolor reporters are a powerful platform for simultaneously tracking discrete cell populations. Here, combining Dre-rox and Cre-loxP systems, we generated a new dual-recombinase reporter system, called Rosa26 traffic light reporter (R26-TLR), to monitor red, green, and yellow fluorescence. Using this new reporter system with the three distinct fluorescent reporters combined on one allele, we found that the readouts of the two recombinases Cre and Dre simultaneously reflect Cre+Dre-, Cre-Dre+, and Cre+Dre+ cell lineages. As proof of principle, we show specific labeling in three distinct progenitor/stem cell populations, including club cells, AT2 cells, and bronchoalveolar stem cells, in Sftpc-DreER;Scgb1a1-CreER;R26-TLR mice. By using this new dual-recombinase reporter system, we simultaneously traced the cell fate of these three distinct cell populations during lung repair and regeneration, providing a more comprehensive picture of stem cell function in distal airway repair and regeneration. We propose that this new reporter system will advance developmental and regenerative research by facilitating a more sophisticated genetic approach to studying in vivo cell fate plasticity.

中文翻译:

双报告子在单个等位基因上追踪三细胞谱系。

遗传谱系追踪被广泛用于研究器官发育和组织再生。多色报道分子是一个强大的平台,可同时跟踪离散的细胞群。在这里,结合了Dre-rox和Cre-loxP系统,我们生成了一个新的双重重组报告基因系统,称为Rosa26红绿灯报告基因(R26-TLR),用于监视红色,绿色和黄色荧光。使用这种新的报告基因系统,将三个不同的荧光报告基因结合在一个等位基因上,我们发现两个重组酶Cre和Dre的读数同时反映了Cre + Dre-,Cre-Dre +和Cre + Dre +细胞谱系。作为原理证明,我们在Sftpc-DreER; Scgb1a1-CreER; R26-TLR小鼠中显示了三个不同的祖细胞/干细胞群体中的特异性标记,包括俱乐部细胞,AT2细胞和支气管肺泡干细胞。通过使用这种新的双重重组酶报告系统,我们在肺修复和再生期间同时追踪了这三个不同细胞群的细胞命运,从而提供了干细胞在远端气道修复和再生中功能的更全面的描述。我们建议,这个新的报告系统将通过促进更复杂的遗传方法来研究体内细胞命运可塑性来促进发展和再生研究。
更新日期:2020-01-17
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