Decoder-seq enhances mRNA capture efficiency in spatial RNA sequencing

Nat Biotechnol. 2024 Nov;42(11):1735-1746. doi: 10.1038/s41587-023-02086-y. Epub 2024 Jan 16.

Abstract

Spatial transcriptomics technologies with high resolution often lack high sensitivity in mRNA detection. Here we report a dendrimeric DNA coordinate barcoding design for spatial RNA sequencing (Decoder-seq), which offers both high sensitivity and high resolution. Decoder-seq combines dendrimeric nanosubstrates with microfluidic coordinate barcoding to generate spatial arrays with a DNA density approximately ten times higher than previously reported methods while maintaining flexibility in resolution. We show that the high RNA capture efficiency of Decoder-seq improved the detection of lowly expressed olfactory receptor (Olfr) genes in mouse olfactory bulbs and contributed to the discovery of a unique layer enrichment pattern for two Olfr genes. The near-cellular resolution provided by Decoder-seq has enabled the construction of a spatial single-cell atlas of the mouse hippocampus, revealing dendrite-enriched mRNAs in neurons. When applying Decoder-seq to human renal cell carcinomas, we dissected the heterogeneous tumor microenvironment across different cancer subtypes and identified spatial gradient-expressed genes related to epithelial-mesenchymal transition with the potential to predict tumor prognosis and progression.

MeSH terms

  • Animals
  • Carcinoma, Renal Cell / genetics
  • Carcinoma, Renal Cell / pathology
  • Gene Expression Profiling / methods
  • Humans
  • Kidney Neoplasms / genetics
  • Kidney Neoplasms / pathology
  • Mice
  • Olfactory Bulb / metabolism
  • RNA, Messenger* / genetics
  • RNA, Messenger* / metabolism
  • Receptors, Odorant / genetics
  • Receptors, Odorant / metabolism
  • Sequence Analysis, RNA* / methods
  • Single-Cell Analysis / methods
  • Tumor Microenvironment

Substances

  • RNA, Messenger
  • Receptors, Odorant