实时体内RNA成像可以增进我们对生理病理过程的理解。然而，大多数以核酸为基础的传感器对核酸酶的抵抗力较差，光物理特性有限，因此不太适用于这个目的。为了解决这个问题，我们开发了一种半导体聚合物纳米核酸探针（SENSE），用于在活体小鼠中进行肿瘤免疫转录组成像。SENSE由一个与识别DNA链共价连接的半导体聚合物骨架和一个染料标记的信号DNA链组成。当探针探测到目标T淋巴细胞的转录物（Gzmb：髓外酶B）时，信号链会从SENSE释放出来，导致信号染料荧光增强，并具有与转录水平的信号相关性的超灵敏度。SENSE中SP核的持续发光也用作跟踪肿瘤中SENSE的吸收的内部参考。因此，SENSE具有双信号通道，可以对活体小鼠肿瘤中的Gzmb转录物进行比色成像，以评估化疗免疫疗法。此外，它已经展示了与流式细胞术和qPCR相当的敏感性和特异性，还提供了一种更快速、更简单的检测切除肿瘤中T细胞的方法。因此，SENSE代表了一种有前景的体内RNA成像工具。本文受版权保护。保留所有权利。

Real-time in vivo imaging of RNA can enhance our understanding of physio-pathological processes. However, most nucleic acid-based sensors have poor resistance to nucleases and limited photophysical properties, making them suboptimal for this purpose. To address this, we have developed a semiconducting polymer nanospherical nucleic acid probe (SENSE) for transcriptomic imaging of cancer immunity in living mice. SENSE consists of a semiconducting polymer backbone covalently linked with a recognition DNA strand, which is complemented by a dye-labelled signal DNA strand. Upon detection of a targeted T lymphocyte transcript (Gzmb: granzyme B), the signal strand is released from SENSE, leading to a fluorescence enhancement of the signal dye and superb sensitivity for signal correlation with transcript levels. The always-on fluorescence of the SP core in SENSE also serves as an internal reference for tracking SENSE uptake in tumours. Thus, SENSE has the du-al-signal channel that allows for ratiometric imaging of Gzmb transcripts in the tumour of living mice for evaluating chemo-immunotherapy; moreover, it has demonstrated sensitivity and specificity comparable to flow cytometry and qPCR, with the added benefit of a faster and simpler means of T cell detection in resected tumours. Therefore, SENSE represents a promising tool for in vivo RNA imaging. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.