Abstract:
Single-cell multi-omics can provide a unique perspective on tumor cellular heterogeneity. Most previous single-cell-whole-genome-RNA-sequencing (scWGS-RNA-seq) methods demonstrate utility with intact cells from fresh samples. Among them, most are not applicable to frozen samples that cannot generate intact single-cell suspensions. We have developed scONE-seq, a versatile scWGS-RNA-seq method that amplifies single-cell DNA and RNA without separating them from each other. Compared with existing methods, scONE-seq is a one-tube reaction that eliminates loss due to DNA and RNA separation and thus is compatible with frozen biobanked tissue. We applied it to analyze a 2-year-frozen astrocytoma sample and identified a unique transcriptionally normal-like tumor clone. scONE-seq makes it possible to perform single-cell multi-omics interrogation on the vast quantities of biobanked tissue, to this end we have also developed a droplet array platform for implementing multi-step single-cell reactions at scale.