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| Item Type: | Preprint |
|---|---|
| Title: | Transcript-specific enrichment enables profiling rare cell states via scRNA-seq |
| Creators Name: | Abay, T., Stickels, R.R., Takizawa, M.T., Nalbant, B.N., Hsieh, Y.H., Hwang, S., Snopkowski, C., Yu, K.K.H., Abou-Mrad, Z., Tabar, V., Ludwig, L.S., Chaligne, R., Satpathy, A.T. and Lareau, C.A. |
| Abstract: | Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing (scRNA-seq) identifies many rare populations of interest that express specific marker transcript combinations, traditional flow sorting limits our ability to enrich these populations for further profiling, including requiring cell surface markers with high-fidelity antibodies. Additionally, 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. To address these limitations, we describe Programmable Enrichment via RNA Flow-FISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations from complex cellular mixtures defined by the presence or absence of specific RNA transcripts. Across immune populations (n = 141,227 cells) and fresh-frozen and formalin-fixed paraffin-embedded brain tissue (n = 29,522 nuclei), we demonstrate the sorting logic that can be used to enrich for cell populations via RNA-based cytometry followed by high-throughput scRNA-seq. Our approach provides a rational, programmable method for studying rare populations identified by one or more marker transcripts. |
| Source: | bioRxiv |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Article Number: | 2024.03.27.587039 |
| Date: | 27 March 2024 |
| Official Publication: | https://doi.org/10.1101/2024.03.27.587039 |
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