基于多色编码的多路复用、空间分辨成像揭示了单细胞RNA表观遗传修饰的异质性。
Multichrome encoding-based multiplexed, spatially resolved imaging reveals single-cell RNA epigenetic modifications heterogeneity.
作者信息
Mao Dongsheng, Tang Xiaochen, Zhang Runchi, Hu Song, Gou Hongquan, Zhang Penghui, Li Wenxing, Pan Qiuhui, Shen Bing, Zhu Xiaoli
机构信息
Shanghai Tenth People's Hospital of Tongji University, Shanghai, PR China.
Department of Clinical Laboratory Medicine, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China.
出版信息
Nat Commun. 2025 Jan 22;16(1):958. doi: 10.1038/s41467-025-56331-w.
Understanding the heterogeneity of epigenetic modifications within single cells is pivotal for unraveling the nature of the complexity of gene expression and cellular function. In this study, we have developed a strategy based on multichrome encoding and "AND" Boolean logic recognition for multiplexed, spatially resolved imaging of single-cell RNA epigenetic modifications, termed as PRoximity Exchange-assisted Encoding of Multichrome (PREEM). Through the implementation of this strategy, we can now map the expression and nuclear distribution of multiple site-specific RNA N6-methyladenosine (mA) modifications at the single-molecule resolution level in single-cells, and reveal the previously unknown heterogeneity. Notably, we demonstrate how these patterns change after treatment with various drugs. Moreover, cyclic imaging with tailed DNA self-assembly further suggest the scalability and adaptability of PREEM's design. As an innovative epigenetic modification imaging tool, PREEM not only broadens the horizons of single-cell epigenetics research, enabling joint analysis of multiple targets beyond the limitations of imaging channels, but also reveals cell-to-cell variability, thereby enhancing our capacity to explore cellular functions.
了解单细胞内表观遗传修饰的异质性对于揭示基因表达和细胞功能复杂性的本质至关重要。在本研究中,我们开发了一种基于多色编码和“与”布尔逻辑识别的策略,用于单细胞RNA表观遗传修饰的多重、空间分辨成像,称为多色邻近交换辅助编码(PREEM)。通过实施该策略,我们现在可以在单细胞中以单分子分辨率水平绘制多个位点特异性RNA N6-甲基腺苷(m6A)修饰的表达和核分布,并揭示以前未知的异质性。值得注意的是,我们展示了用各种药物处理后这些模式如何变化。此外,使用带尾DNA自组装的循环成像进一步表明了PREEM设计的可扩展性和适应性。作为一种创新的表观遗传修饰成像工具,PREEM不仅拓宽了单细胞表观遗传学研究的视野,能够在成像通道限制之外对多个靶点进行联合分析,还揭示了细胞间的变异性,从而增强了我们探索细胞功能的能力。