Development Of Single Cell Holo-Transcriptome Sequencing Technology And Application In Liver Cancer

There are no two cells exactly same at transcriptional level.Regular bulk RNA-sequencing requires tens of thousands of cells and provides an average view of gene expression,which masks the cell-to-cell variation and heterogeneity.Single cell sequencing holds great potential for exploring biological systems with unprecedented resolution.In recent years,the single cell RNA-seq technologies have been develop rapidly and serve as a powerful approach to dissect cellular heterogeneity and identify rare cell types in various research,such as early embryonic development to tissue and organ development,immunology and oncology.However,pre-amplification of large cDNAs may cause severe quantitative errors,losses of full-length strand information and the inability of simultaneously observing small RNA and mRNA dual transcriptomes.We have developed a single cell holo-transcriptome sequencing method(Holo-Seq)that overcomes all above hurdles.Holo-Seq has the same quantitative accuracy and full-length transcript coverage as bulk RNA-Seq.Most importantly,Holo-Seq can profile full-length strand information and enables the simultaneous observation of small RNA and mRNA transcriptomes in a single cell.Cellular heterogeneity is a fundamental characteristic of many cancers.To elucidate the cell type composition of a tumor and explore potential markers that will be useful to diagnostics,therapeutics and prognostics,we applied Holo-seq to single cells isolated from human hepatocellular carcinoma and put efforts to defining cell identities based on a molecular profiling.We also observed the genome-wide super-enhancer activity and hepatic neoplasm kinetics of these HCC cells.We expect Holo-seq to enter clinics to facilitate more personalized therapeutic decisions for patients in near future.