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<p>Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing identifies rare populations that express specific marker transcript combinations, traditional flow sorting requires cell surface markers with high-fidelity antibodies, limiting our ability to interrogate these populations. In addition, many single-cell studies require the isolation of nuclei from tissue, eliminating the ability to enrich learned rare cell states based on extranuclear protein markers. In the present report, we addressed these limitations by developing Programmable Enrichment via RNA FlowFISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations defined by the abundance of specific RNA transcripts. Across immune populations (<i>n</i> = 184,126 cells) and fresh-frozen and formalin-fixed, paraffin-embedded brain tissue (<i>n</i> = 33,145 nuclei), we demonstrated that programmable sorting logic via RNA-based cytometry can isolate rare cell populations and uncover phenotypic heterogeneity via downstream, high-throughput, single-cell genomics analyses.</p>

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Transcript-specific enrichment enables profiling of rare cell states via single-cell RNA sequencing

  • 期刊论文

  • 发表日期：

    2025年1月8日

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  • Tsion Abay,
  • Robert R. Stickels,
  • Meril T. Takizawa,
  • Benan N. Nalbant,
  • Yu-Hsin Hsieh,
  • Sidney Hwang,
  • Catherine Snopkowski,
  • Kenny Kwok Hei Yu,
  • Zaki Abou-Mrad,
  • Viviane Tabar,
  • Brooke E. Howitt,
  • Leif S. Ludwig,
  • Ronan Chaligné,
  • Ansuman T. Satpathy,
  • Caleb A. Lareau

摘要

Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing identifies rare populations that express specific marker transcript combinations, traditional flow sorting requires cell surface markers with high-fidelity antibodies, limiting our ability to interrogate these populations. In addition, many single-cell studies require the isolation of nuclei from tissue, eliminating the ability to enrich learned rare cell states based on extranuclear protein markers. In the present report, we addressed these limitations by developing Programmable Enrichment via RNA FlowFISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations defined by the abundance of specific RNA transcripts. Across immune populations (n = 184,126 cells) and fresh-frozen and formalin-fixed, paraffin-embedded brain tissue (n = 33,145 nuclei), we demonstrated that programmable sorting logic via RNA-based cytometry can isolate rare cell populations and uncover phenotypic heterogeneity via downstream, high-throughput, single-cell genomics analyses.