Single-Molecule Real-Time (SMRT) Full-Length RNA-Sequencing Reveals Novel and Distinct mRNA Isoforms in Human Bone Marrow Cell Subpopulations.
作者： Anton Wellstein , Anne Deslattes Mays , Marcel Schmidt , Garrett Graham , Elizabeth Tseng
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摘要: Hematopoietic cells are continuously replenished from progenitor that reside in the bone marrow. To evaluate molecular changes during this process, we analyzed transcriptomes of freshly harvested human marrow (lineage-negative) and differentiated (lineage-positive) by single-molecule real-time (SMRT) full-length RNA-sequencing. This analysis revealed a ~5-fold higher number transcript isoforms than previously detected showed distinct composition individual characteristic for subpopulations. A detailed messenger RNA (mRNA) transcribed ANXA1 EEF1A1 loci confirmed their composition. The expression proteins predicted transcriptome was evaluated mass spectrometry validated unknown protein e.g., EEF1A1. These distinguished lineage negative cell population positive population. Finally, expressed paralogous gene (e.g., CFD, GATA2, HLA-A, B, C) also subpopulations but were only detectable sequencing. Thus, qualitatively genomic separate indicating complex transcriptional regulation isoform generation hematopoiesis.
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