Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK.
Development. 2014 Apr;141(7):1589-98. doi: 10.1242/dev.105254.
Cell lineage analysis enables us to address pivotal questions relating to: the embryonic origin of cells and sibling cell relationships in the adult body; the contribution of progenitors activated after trauma or disease; and the comparison across species in evolutionary biology. To address such fundamental questions, several techniques for clonal labelling have been developed, each with its shortcomings. Here, we report a novel method, CLoNe that is designed to work in all vertebrate species and tissues. CLoNe uses a cocktail of labelling, targeting and transposition vectors that enables targeting of specific subpopulations of progenitor types with a combination of fluorophores resulting in multifluorescence that describes multiple clones per specimen. Furthermore, transposition into the genome ensures the longevity of cell labelling. We demonstrate the robustness of this technique in mouse and chick forebrain development, and show evidence that CLoNe will be broadly applicable to study clonal relationships in different tissues and species.
细胞的胚胎起源和成年体内同系细胞的关系；创伤或疾病后激活的祖细胞的贡献；以及进化生物学中不同物种间的比较。为了解决这些基本问题，已经开发了几种克隆标记技术，但每种技术都有其缺点。在这里，我们报告了一种新的方法 CLoNe，该方法旨在适用于所有脊椎动物物种和组织。CLoNe 使用标记、靶向和转座子的混合物，能够针对特定类型的祖细胞亚群进行靶向，并用荧光团组合进行标记，从而对每个标本进行多重荧光标记，描述多个克隆。此外，转座到基因组中确保了细胞标记的持久性。我们在小鼠和鸡前脑发育中证明了该技术的稳健性，并提供了证据表明 CLoNe 将广泛适用于研究不同组织和物种中的克隆关系。
