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High-throughput single-cell proteomics and transcriptomics from same cells with a nanoliter-scale, spin-transfer approach.

Pranav Dawar·Lye Meng Markillie·Sarah M Williams·Hugh D Mitchell·Johannes W Bagnoli·Joshua Cantlon-Bruce·Anjali Seth·Carter C Bracken·Ljiljana Paša-Tolić·Ying Zhu·James M Fulcher
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Abstract

Single-cell multiomic platforms provide a comprehensive snapshot of cellular states and cell types by offering critical insights into the spatiotemporal regulation of biomolecular networks at a systems level, thereby defining the basis of multicellularity. Here, we introduce nanoSPINS, an advanced platform that enables high-throughput profiling and integrative analysis of the transcriptome and proteome from the same single cells using RNA sequencing and isobaric labeling LC-MS-based proteomics, respectively. NanoSPINS can efficiently transfer mRNA-containing droplets across two microarrays via a centrifugation-based approach, while proteins are retained on the initial platform. Benchmarking of nanoSPINS on two cell lines demonstrates its ability to generate global proteomic and transcriptomic profiles that align well with previously established methodologies/platforms. The incorporation of isobaric TMTpro labeling into this single-cell multiomics platform significantly enhances the throughput of single-cell proteomic analyses. Through the high-throughput quantification of the proteome and transcriptome, nanoSPINS not only facilitates the identification of molecular features at both mRNA and protein level but also provides larger sample sizes for improved statistical power in clustering and differential abundance. Given the broad applicability of single-cell multiomics in biological research and clinical settings, we believe nanoSPINS represents a powerful platform for the characterization of heterogeneous cell populations.

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Single-Cell AnalysisProteomicsHumansSingle-Cell Gene Expression AnalysisTranscriptomeMultiomicsProteomeNanotechnologyGene Expression ProfilingJournal ArticleResearch Support, Non-U.S. Gov't

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Pranav Dawar, Lye Meng Markillie, Sarah M Williams, Hugh D Mitchell, Johannes W Bagnoli, Joshua Cantlon-Bruce, Anjali Seth, Carter C Bracken, Ljiljana Paša-Tolić, Ying Zhu, James M Fulcher. 2026-03-31. High-throughput single-cell proteomics and transcriptomics from same cells with a nanoliter-scale, spin-transfer approach.. https://doi.org/10.1039/d5lc01008j

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