Reference long-read isoform-aware transcriptomes of 4 human peripheral blood lymphocyte subsets

Area of Science:

• Immunology
• Genomics
• Molecular Biology

Background:

• Long-read sequencing offers accurate full-length mRNA transcript isoform analysis.
• Investigating lymphocyte functional plasticity is crucial for understanding human health and disease.
• A lack of long-read, isoform-aware reference transcriptomes for human lymphocytes hinders research.

Purpose of the Study:

• To create comprehensive, long-read reference transcriptomes for human circulating lymphocyte populations.
• To provide essential benchmarks for transcriptomic studies involving lymphocytes.
• To support research into lymphocyte functional plasticity and its role in health and disease.

Main Methods:

• Purification of four human lymphocyte populations (CD4+ T, CD8+ T, NK, Pan B cells) from peripheral blood.
• High-quality RNA extraction (RIN > 8) for downstream analyses.
• Isoform sequencing and parallel RNA-Sequencing (RNA-Seq) for transcriptomic profiling.

Main Results:

• Identification of numerous novel polyadenylated transcript isoforms across all analyzed lymphocyte populations.
• Validation of novel isoforms using complementary data from both isoform sequencing and RNA-Seq.
• Generation of high-quality datasets meeting rigorous quality metrics.

Conclusions:

• The developed reference transcriptomes provide a valuable resource for the scientific community.
• These datasets will significantly advance the study of human circulating lymphocytes.
• The findings facilitate future research into lymphocyte biology, functional plasticity, and associated diseases.