State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China.
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Nat Biomed Eng. 2024 Jul;8(7):872-889. doi: 10.1038/s41551-024-01205-7. Epub 2024 May 14.
Technology for spatial multi-omics aids the discovery of new insights into cellular functions and disease mechanisms. Here we report the development and applicability of multi-omics in situ pairwise sequencing (MiP-seq), a method for the simultaneous detection of DNAs, RNAs, proteins and biomolecules at subcellular resolution. Compared with other in situ sequencing methods, MiP-seq enhances decoding capacity and reduces sequencing and imaging costs while maintaining the efficacy of detection of gene mutations, allele-specific expression and RNA modifications. MiP-seq can be integrated with in vivo calcium imaging and Raman imaging, which enabled us to generate a spatial multi-omics atlas of mouse brain tissues and to correlate gene expression with neuronal activity and cellular biochemical fingerprints. We also report a sequential dilution strategy for resolving optically crowded signals during in situ sequencing. High-throughput in situ pairwise sequencing may facilitate the multidimensional analysis of molecular and functional maps of tissues.
空间多组学技术有助于深入了解细胞功能和疾病机制。本文报告了多组学原位两两测序（MiP-seq）的开发和适用性，该方法可在亚细胞分辨率下同时检测 DNA、RNA、蛋白质和生物分子。与其他原位测序方法相比，MiP-seq 提高了解码能力，降低了测序和成像成本，同时保持了基因突变、等位基因特异性表达和 RNA 修饰的检测效果。MiP-seq 可与体内钙成像和拉曼成像相结合，使我们能够生成小鼠脑组织的空间多组学图谱，并将基因表达与神经元活动和细胞生化指纹相关联。我们还报告了一种用于解决原位测序过程中光学拥挤信号的顺序稀释策略。高通量原位两两测序可能有助于组织的分子和功能图谱的多维分析。
