Accurately characterizing cell types within complex cell structures provides invaluable information for comprehending the cellular status during biological processes. In this study, we have developed an miRNA-switch cocktail platform capable of reporting and tracking the activities of multiple miRNAs (microRNAs) at the single-cell level, while minimizing disruption to the cell culture. Drawing on the principles of traditional miRNA-sensing mRNA switches, our platform incorporates subcellular tags and employs intelligent engineering to segment three subcellular regions using two fluorescent proteins. These designs enable the quantification of multiple miRNAs within the same cell. Through our experiments, we have demonstrated the platform’s ability to track marker miRNA levels during cell differentiation and provide spatial information of heterogeneity on outlier cells exhibiting extreme miRNA levels. Importantly, this platform offers real-time and miRNA reporting, allowing for multidimensional evaluation of cell profile and paving the way for a comprehensive understanding of cellular events during biological processes.

中文翻译：

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用于活细胞实时分析的多重 miRNA 报告平台

准确表征复杂细胞结构中的细胞类型为理解生物过程中的细胞状态提供了宝贵的信息。在这项研究中，我们开发了一个 miRNA 开关鸡尾酒平台，能够在单细胞水平上报告和跟踪多个 miRNA (microRNA) 的活动，同时最大限度地减少对细胞培养的破坏。我们的平台借鉴传统 miRNA 传感 mRNA 开关的原理，结合了亚细胞标签，并采用智能工程，使用两种荧光蛋白分割三个亚细胞区域。这些设计能够对同一细胞内的多个 miRNA 进行定量。通过我们的实验，我们证明了该平台能够在细胞分化过程中跟踪标记 miRNA 水平，并提供表现出极端 miRNA 水平的异常细胞的异质性空间信息。重要的是，该平台提供实时和 miRNA 报告，允许对细胞概况进行多维评估，并为全面了解生物过程中的细胞事件铺平道路。