Single-cell isoform analysis in human immune cells

Genome Biol. 2022 Feb 7;23(1):47. doi: 10.1186/s13059-022-02615-z.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Profiling / methods
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Leukocytes, Mononuclear*
  • Protein Isoforms / genetics
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis*

Substances

  • Protein Isoforms