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Ultra-high Throughput Single-cell Analysis Of Proteins And RNAs By Split-pool Synthesis

Maeve O’Huallachain, Felice-Alessio Bava, Mary Shen, Carolina Dallett, Sri Paladugu, Nikolay Samusik, Simon Yu, Razika Hussein, Grantland R. Hillman, Samuel Higgins, Melanie Lou, Angelica Trejo, Laura Qin, Yu Chuan Tai, Shigemi M. Kinoshita, Astraea Jager, Deval Lashkari, Yury Goltsev, Sedide Ozturk, Garry P. Nolan

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AbstractSingle-cell omics provide insight into cellular heterogeneity and function. Recent technological advances have accelerated single-cell analyses, but workflows remain expensive and complex. We present a method enabling simultaneous, ultra-high throughput single-cell barcoding of millions of cells for targeted analysis of proteins and RNAs. Quantum barcoding (QBC) avoids isolation of single cells by building cell-specific oligo barcodes dynamically within each cell. With minimal instrumentation (four 96-well plates and a multichannel pipette), cell-specific codes are added to each tagged molecule within cells through sequential rounds of classical split-pool synthesis. Here we show the utility of this technology in mouse and human model systems for as many as 50 antibodies to targeted proteins and, separately, >70 targeted RNA regions. We demonstrate that this method can be applied to multi-modal protein and RNA analyses. It can be scaled by expansion of the split-pool process and effectively renders sequencing instruments as versatile multi-parameter flow cytometers.