Single-cell isoform analysis in human immune cells
Roger Volden, Christopher Vollmers
Abstract
High-throughput single-cell analysis today is facilitated by protocols like the 10X Genomics platform or Drop-Seq which generate cDNA pools in which the origin of a transcript is encoded at its 5' or 3' end. Here, we used R2C2 to sequence and demultiplex 12 million full-length cDNA molecules generated by the 10X Genomics platform from ~3000 peripheral blood mononuclear cells. We use these reads, independent from Illumina data, to identify B cell, T cell, and monocyte clusters and generate isoform-level transcriptomes for cells and cell types. Finally, we extract paired adaptive immune receptor sequences unique to each T and B cell.
Topics & Concepts
BiologyTranscriptomeComplementary DNAComputational biologyGene isoformGenomicsPeripheral blood mononuclear cellSingle-cell analysisImmune systemFunctional genomicsCellGeneticsMolecular biologyGeneGene expressionGenomeIn vitroSingle-cell and spatial transcriptomicsT-cell and B-cell ImmunologyImmune Cell Function and Interaction