PUFFFIN:用于标记细胞邻域的超亮、可定制的单质粒系统。

IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY EMBO Journal Pub Date : 2024-09-01 Epub Date: 2024-07-12 DOI:10.1038/s44318-024-00154-w
Tamina Lebek, Mattias Malaguti, Giulia Lm Boezio, Lida Zoupi, James Briscoe, Alistair Elfick, Sally Lowell
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引用次数: 0

摘要

细胞通讯可协调发育过程、维持体内平衡并导致疾病。因此,了解共享环境中细胞之间的关系至关重要。在这里,我们介绍了一种基于正超电荷荧光蛋白 s36GFP 的分泌和吸收的细胞邻居标记系统--正超亮荧光融合识别邻居(PUFFFIN)。我们将 s36GFP 与 mNeonGreen 或 HaloTag 融合,从而实现了超亮、灵敏、颜色可选的标记。分泌细胞将 PUFFFIN 传递给邻近细胞,同时保留核 mCherry,这样就能直接识别、分离和研究活的邻近细胞。PUFFFIN 可以通过一个可定制的单质粒构建体输送到细胞、组织或胚胎中,该构建体由可互换的组件组成,并可选择加入任何转基因。这种多功能性使我们能够在细胞培养或体内操纵细胞特性,同时标记周围的细胞。我们利用 PUFFFIN 来探究多能细胞在脱离幼稚多能性过程中是否会调整分化速度,以便与周围细胞同步。PUFFFIN 提供了一种简单、灵敏、可定制的方法,用于分析细胞对自然或诱导的细胞特性或行为变化的非自主反应。
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PUFFFIN: an ultra-bright, customisable, single-plasmid system for labelling cell neighbourhoods.

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.

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来源期刊
EMBO Journal
EMBO Journal 生物-生化与分子生物学
CiteScore
18.90
自引率
0.90%
发文量
246
审稿时长
1.5 months
期刊介绍: The EMBO Journal has stood as EMBO's flagship publication since its inception in 1982. Renowned for its international reputation in quality and originality, the journal spans all facets of molecular biology. It serves as a platform for papers elucidating original research of broad general interest in molecular and cell biology, with a distinct focus on molecular mechanisms and physiological relevance. With a commitment to promoting articles reporting novel findings of broad biological significance, The EMBO Journal stands as a key contributor to advancing the field of molecular biology.
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