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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

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

Patricia Hayes Doyle1,2, Madeline L Page1,2,3, J Anthony Brandon1,2,3

  • 1Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY, United States.

Frontiers in Genetics
|June 12, 2026
PubMed
Summary
This summary is machine-generated.

Long-read single-cell sequencing reveals novel gene isoforms in human immune cells, including megakaryocytes. This advanced technique maps the full isoform landscape, enhancing our understanding of cellular diversity and disease mechanisms.

Keywords:
Oxford Nanopore Technologies (ONT)PBMCsPIPseqRNA isoformsalternative splicingisoform discoverylong-read single-cell RNA sequencing

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Area of Science:

  • Genomics
  • Molecular Biology
  • Immunology

Background:

  • Long-read single-cell RNA sequencing offers unprecedented resolution for studying human health and disease.
  • Current methods struggle to capture isoform-level diversity and define cell types comprehensively.

Purpose of the Study:

  • To generate the largest long-read single-cell dataset of human peripheral blood mononuclear cells (PBMCs) to date.
  • To profile isoform usage across immune cells and identify novel isoforms.
  • To investigate cell-type-specific isoform expression patterns and their functional implications.

Main Methods:

  • Utilized a modified, microfluidic-free PIPseq workflow.
  • Adapted a computational pipeline for Oxford Nanopore long-read sequencing.
  • Generated and analyzed a large-scale long-read single-cell PBMC dataset.

Main Results:

  • Identified 126 novel isoforms from known and new genes with cell-type-specific patterns.
  • Characterized marker gene isoform expression across various immune cell types.
  • Discovered non-canonical variants of GZMB and CD3G enriched in unexpected cell types, including megakaryocytes.

Conclusions:

  • Long-read single-cell sequencing is versatile for mapping the isoform landscape (isonome) in biological contexts.
  • This approach enables the integration of isoform signatures within the functional context of cellular biology.
  • The study expands the utility of long-read single-cell technologies for discovering novel transcripts and understanding cellular diversity.