State Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing, Jiangsu, China.
Department of Neurology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, Jiangsu, China.
PLoS One. 2024 Jul 25;19(7):e0305977. doi: 10.1371/journal.pone.0305977. eCollection 2024.
Profiling gene expression while preserving cell locations aids in the comprehensive understanding of cell fates in multicellular organisms. However, simple and flexible isolation of microregions of interest (mROIs) for spatial transcriptomics is still challenging. We present a laser-induced forward transfer (LIFT)-based method combined with a full-length mRNA-sequencing protocol (LIFT-seq) for profiling region-specific tissues. LIFT-seq demonstrated that mROIs from two adjacent sections could reliably and sensitively detect and display gene expression. In addition, LIFT-seq can identify region-specific mROIs in the mouse cortex and hippocampus. Finally, LIFT-seq identified marker genes in different layers of the cortex with very similar expression patterns. These genes were then validated using in situ hybridization (ISH) results. Therefore, LIFT-seq will be a valuable and efficient technique for profiling the spatial transcriptome in various tissues.
在保留细胞位置的同时进行基因表达谱分析有助于全面了解多细胞生物中的细胞命运。然而，简单灵活地分离用于空间转录组学的微区（mROI）仍然具有挑战性。我们提出了一种基于激光诱导正向转移（LIFT）的方法，结合全长 mRNA 测序方案（LIFT-seq），用于分析区域特异性组织。LIFT-seq 表明，来自两个相邻切片的 mROI 可以可靠且灵敏地检测和显示基因表达。此外，LIFT-seq 可以识别小鼠皮层和海马体中的区域特异性 mROI。最后，LIFT-seq 鉴定了皮层不同层中具有非常相似表达模式的标记基因。然后使用原位杂交（ISH）结果对这些基因进行验证。因此，LIFT-seq 将是一种非常有价值和有效的技术，可用于分析各种组织中的空间转录组。
