Hu Senhao, Wang Wenjing, Zou Yu, Li Chunmei, Zou Hongyan, Huang Chengzhi, Zhan Lei
Key Laboratory of Biomedical Analytics, Chongqing Science and Technology Bureau, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
Molecules. 2025 Aug 29;30(17):3551. doi: 10.3390/molecules30173551.
The development of low-background, facile, and robust fluorescent nanoprobes for imaging and monitoring of intracellular mRNA changes remains a great challenge. Taking advantage of the high fluorescence quenching efficiency of core-shell gold@polydopamine (Au@PDA) nanocomposites and Ca-promoting DNA adsorption stability, a simple and universal bioconjugate strategy was designed to a construct fluorescent nanoprobe for highly efficient tumor-related mRNA imaging. The fluorescence of Cy5-labeled DNA was quenched up to 92.38% by the AuNP and PDA via nanometal surface energy transfer (NSET) and photoinduced electron transfer (PET), respectively. TK1 mRNA, a biomarker of tumor growth, initiates hybridization and results in fluorescence recovery, which built the foundation for identifying the expression level changes in living cells. More importantly, three kinds of tumor-related mRNA (TK1 mRNA, GalNAc-T mRNA, and C-myc mRNA) can be detected simultaneously with different fluorophore-modified recognition sequences, which can avoid false positive signals and improve the reliability of cancer diagnostic, holding great promise for cancer diagnosis, prognosis, and therapy.
开发用于成像和监测细胞内mRNA变化的低背景、简便且稳健的荧光纳米探针仍然是一项巨大挑战。利用核壳金@聚多巴胺（Au@PDA）纳米复合材料的高荧光猝灭效率以及钙促进DNA吸附稳定性，设计了一种简单通用的生物共轭策略来构建用于高效肿瘤相关mRNA成像的荧光纳米探针。通过纳米金属表面能量转移（NSET）和光致电子转移（PET），AuNP和PDA分别将Cy5标记的DNA的荧光猝灭高达92.38%。肿瘤生长的生物标志物胸苷激酶1（TK1）mRNA引发杂交并导致荧光恢复，这为识别活细胞中的表达水平变化奠定了基础。更重要的是，三种肿瘤相关mRNA（TK1 mRNA、N-乙酰半乳糖胺转移酶（GalNAc-T）mRNA和C- myc mRNA）可以用不同荧光团修饰的识别序列同时检测，这可以避免假阳性信号并提高癌症诊断的可靠性，在癌症诊断、预后和治疗方面具有巨大潜力。
