A light-gated transcriptional recorder for detecting cell-cell contacts.
A light-gated transcriptional recorder for detecting cell-cell contacts.
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DOI：
10.7554/elife.70881
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发表时间：
2022-03-21
期刊：
影响因子：
7.7
通讯作者：
Ting AY
中科院分区：
文献类型：
--
作者：
Cho KF;Gillespie SM;Kalogriopoulos NA;Quezada MA;Jacko M;Monje M;Ting AY
检测细胞间接触的技术是研究广泛生物学过程的有力工具，从神经元信号传导到肿瘤微环境中的癌症-免疫相互作用。在这里，我们报告TRACC（细胞-细胞接触激活的转录读出），一种基于GPCR的细胞接触转录记录器，它将接触事件转化为稳定的转基因表达。TRACC源自我们先前的蛋白质-蛋白质相互作用记录器SPARK（Kim等人，2017）和SPARK 2（Kim等人，2019年，在这本杂志上发表。TRACC通过光氧电压感应（LOV）域结合了光门控，提供了用户定义的工具激活时间控制并减少了背景。我们表明，TRACC检测细胞与哺乳动物细胞培养的高特异性和灵敏度的细胞接触，它可以用来询问神经元和神经胶质瘤，一种形式的脑癌之间的相互作用。
Technologies for detecting cell-cell contacts are powerful tools for studying a wide range of biological processes, from neuronal signaling to cancer-immune interactions within the tumor microenvironment. Here, we report TRACC (Transcriptional Readout Activated by Cell-cell Contacts), a GPCR-based transcriptional recorder of cellular contacts, which converts contact events into stable transgene expression. TRACC is derived from our previous protein-protein interaction recorders, SPARK (Kim et al., 2017) and SPARK2 (Kim et al., 2019), reported in this journal. TRACC incorporates light gating via the light-oxygen-voltage-sensing (LOV) domain, which provides user-defined temporal control of tool activation and reduces background. We show that TRACC detects cell-cell contacts with high specificity and sensitivity in mammalian cell culture and that it can be used to interrogate interactions between neurons and glioma, a form of brain cancer.