Abstract:
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.
摘要:
在保留细胞位置的同时分析基因表达有助于全面了解多细胞生物体中的细胞命运。然而,用于空间转录组学的感兴趣的微区域(mROI)的简单和灵活的分离仍然具有挑战性。我们提出了一种基于激光诱导的正向转移(LIFT)的方法,该方法结合了全长mRNA测序方案(LIFT-seq),用于分析区域特异性组织。LIFT-seq证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中的区域特异性mROI。最后,LIFT-seq鉴定了皮质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)结果验证这些基因。因此,LIFT-seq将是一种有价值且有效的技术,用于分析各种组织中的空间转录组。