Abstract：
Cell communication coordinates developmental processes, maintains homeostasis, and contributes to disease. Therefore, understanding the relationship between cells in a shared environment is crucial. Here we introduce Positive Ultra-bright Fluorescent Fusion For Identifying Neighbours (PUFFFIN), a cell neighbour-labelling system based upon secretion and uptake of positively supercharged fluorescent protein s36GFP. We fused s36GFP to mNeonGreen or to a HaloTag, facilitating ultra-bright, sensitive, colour-of-choice labelling. Secretor cells transfer PUFFFIN to neighbours while retaining nuclear mCherry, making identification, isolation, and investigation of live neighbours straightforward. PUFFFIN can be delivered to cells, tissues, or embryos on a customisable single-plasmid construct composed of interchangeable components with the option to incorporate any transgene. This versatility enables the manipulation of cell properties, while simultaneously labelling surrounding cells, in cell culture or in vivo. We use PUFFFIN to ask whether pluripotent cells adjust the pace of differentiation to synchronise with their neighbours during exit from naïve pluripotency. PUFFFIN offers a simple, sensitive, customisable approach to profile non-cell-autonomous responses to natural or induced changes in cell identity or behaviour.
摘要：
细胞通讯协调发育过程，保持稳态，并有助于疾病。因此,理解共享环境中细胞之间的关系至关重要。在这里，我们介绍了用于识别邻居的正超亮荧光融合（PUFFFIN），一种基于分泌和摄取带正电的荧光蛋白s36GFP的细胞邻居标记系统。我们将s36GFP融合到mNeonGreen或HaloTag上，促进超明亮，敏感,颜色的选择标签。保管员细胞将PUFFFIN转移给邻居，同时保留核mCherry，进行识别，隔离,直截了当地调查活着的邻居。PUFFFIN可以传送到细胞，组织，或胚胎上的可定制的单质粒构建体，该构建体由可互换的成分组成，可以选择掺入任何转基因。这种多功能性使细胞属性的操纵，同时标记周围的细胞，在细胞培养或体内。我们使用PUFFFIN来询问多能细胞在从幼稚多能性退出期间是否调整分化速度以与它们的邻居同步。PUFFFIN提供了一个简单的，敏感,可定制的方法来分析细胞对细胞身份或行为的自然或诱导变化的非细胞自主反应。
