Decoding the human PBMC isonome: isoform-level resolution with single-cell long-read transcriptomics.

A modified PIPseq workflow with Oxford Nanopore long-read single-cell RNA sequencing generated the largest human PBMC dataset from one donor, enabling isoform-level detection including megakaryocytes.

• Why it matters: Long-read single-cell RNA sequencing is needed to resolve isoform usage and cellular diversity in health and disease beyond what bulk or short-read gene-level methods can capture.
• What they did: The study profiled isoform usage across immune cells using a microfluidic-free PIPseq workflow and a computational pipeline adapted for Oxford Nanopore sequencing, identifying 126 novel isoforms.
• The result: Novel isoforms showed cell-type-specific patterns, including enriched non-canonical protein-coding variants of GZMB and CD3G in megakaryocytes and monocyte-derived populations, and novel CMC1 and LYAR transcripts with cell-type-specific signatures.