美国哈佛大学Adam E. Cohen研究组取得一项新突破。他们研究出可用于细胞内记录的时间标记磁带。该研究于2023年1月2日发表于国际学术期刊《自然—生物技术》杂志。
研究人员研发出一种工程蛋白纤维，该纤维在其生长过程中结合了不同的荧光标记，以存储类似于自动收报机样的历史记录。嵌入式HaloTag报告器结合了内源性染料，从而产生彩色条纹，将每根光纤的生长映射到相应时间。由感兴趣启动子驱动的eGFP标签共表达记录了转录激活的历史。固定样品上的高分辨率多光谱成像可读取细胞历史，并且相对于HaloTag时间戳的eGFP标记插值可提供准确的绝对计时。
研究人员展示了多西环素诱导HEK细胞转录和培养神经元中cFos启动子活化的记录，在12小时的记录中，单细胞绝对精度为30-40分钟。这种基于蛋白质的自动收报机标签设计可应用到不同生理模式的大规模并行单细胞记录中。
据了解，记录细胞的转录历史将有助于人们更深入地了解细胞的发育轨迹和对外部扰动的反应。
附：英文原文
Title: Time-tagged ticker tapes for intracellular recordings
Author: Lin, Dingchang, Li, Xiuyuan, Moult, Eric, Park, Pojeong, Tang, Benjamin, Shen, Hao, Grimm, Jonathan B., Falco, Natalie, Jia, Bill Z., Baker, David, Lavis, Luke D., Cohen, Adam E.
Issue&Volume: 2023-01-02
Abstract: Recording transcriptional histories of a cell would enable deeper understanding of cellular developmental trajectories and responses to external perturbations. Here we describe an engineered protein fiber that incorporates diverse fluorescent marks during its growth to store a ticker tape-like history. An embedded HaloTag reporter incorporates user-supplied dyes, leading to colored stripes that map the growth of each individual fiber to wall clock time. A co-expressed eGFP tag driven by a promoter of interest records a history of transcriptional activation. High-resolution multi-spectral imaging on fixed samples reads the cellular histories, and interpolation of eGFP marks relative to HaloTag timestamps provides accurate absolute timing. We demonstrate recordings of doxycycline-induced transcription in HEK cells and cFos promoter activation in cultured neurons, with a single-cell absolute accuracy of 30–40minutes over a 12-hour recording. The protein-based ticker tape design we present here could be generalized to achieve massively parallel single-cell recordings of diverse physiological modalities.
DOI: 10.1038/s41587-022-01524-7
Source: https://www.nature.com/articles/s41587-022-01524-7
Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex
本期文章：《自然—生物技术》：Online/在线发表
