Ultra-high-throughput single-cell RNA sequencing and perturbation screening with combinatorial fluidic indexing.
IF: 47.990
Cited by: 92


Cell atlas projects and high-throughput perturbation screens require single-cell sequencing at a scale that is challenging with current technology. To enable cost-effective single-cell sequencing for millions of individual cells, we developed 'single-cell combinatorial fluidic indexing' (scifi). The scifi-RNA-seq assay combines one-step combinatorial preindexing of entire transcriptomes inside permeabilized cells with subsequent single-cell RNA-seq using microfluidics. Preindexing allows us to load several cells per droplet and computationally demultiplex their individual expression profiles. Thereby, scifi-RNA-seq massively increases the throughput of droplet-based single-cell RNA-seq, and provides a straightforward way of multiplexing thousands of samples in a single experiment. Compared with multiround combinatorial indexing, scifi-RNA-seq provides an easy and efficient workflow. Compared to cell hashing methods, which flag and discard droplets containing more than one cell, scifi-RNA-seq resolves and retains individual transcriptomes from overloaded droplets. We benchmarked scifi-RNA-seq on various human and mouse cell lines, validated it for primary human T cells and applied it in a highly multiplexed CRISPR screen with single-cell transcriptome readout of T cell receptor activation.


Gene Expression

MeSH terms

Cell Line
Clustered Regularly Interspaced Short Palindromic Repeats
Cost-Benefit Analysis
Gene Expression Profiling
High-Throughput Nucleotide Sequencing
Receptors, Antigen, T-Cell
Single-Cell Analysis


Datlinger, Paul
Rendeiro, André F
Boenke, Thorina
Senekowitsch, Martin
Krausgruber, Thomas
Barreca, Daniele
Bock, Christoph

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