This preprint introduces a high-throughput approach to study transfer RNA (tRNA) expression alongside messenger RNA at single-cell resolution. The central aim was to determine how tRNA expression changes as human hematopoietic stem cells are specified into mature blood cell types. The authors sought to map tRNA abundance and splicing across differentiation trajectories and to identify tRNAs associated with stemness, general differentiation, and lineage-specific programs.
The authors present a method they term sc-STM-seq, designed to capture tRNAs and mRNA transcriptomes from the same individual cells. The technique is described as scalable and high-throughput, enabling combined profiling of tRNA species and cellular mRNA expression profiles. The abstract emphasizes that sc-STM-seq allows simultaneous interrogation of both RNA classes in single cells, but specific protocol parameters, library preparation details, and computational processing steps are not reported in the abstract.
sc-STM-seq was applied to human bone marrow samples to profile tRNA expression and splicing across hematopoietic differentiation trajectories. Using this single-cell framework, the authors mapped tRNA patterns across stages of human blood cell development, aligning tRNA expression with known differentiation paths and cell states inferred from mRNA profiles. The abstract does not report sample counts, donor information, or the precise number of cells profiled.
The study reports identification of tRNAs that associate with three broad categories of hematopoietic biology:
These findings indicate that relative tRNA abundance is variable across cell states and may represent a previously underappreciated layer of heterogeneity during hematopoiesis.
Beyond steady-state abundance, the authors also mapped tRNA splicing across differentiation trajectories. The simultaneous measurement of tRNA expression and splicing at single-cell resolution allowed characterization of both quantity and processing state across hematopoietic cell types. The abstract emphasizes that tRNA splicing and expression dynamics were profiled but does not provide quantitative metrics or examples of individual tRNAs or splice variants.
The work culminates in an atlas of the hematopoietic tRNA landscape, presented as a resource for the community. A project web link is provided in the manuscript materials (a bioinformatics URL for the hematopoietic tRNA atlas). The abstract indicates that the atlas and associated analyses are available via that resource, but explicit file types, raw data access, or code availability details are not specified in the abstract.
Competing interests are disclosed for two authors: one author received research funding from several companies and consultancy/honoraria from a list of industry organizations; another author reported licensing fees, research support, honoraria, consultancy roles, and stock ownership. All other authors declared no competing interests. The study lists multiple funders and supporting organizations, including the European Hematology Association, Deutsche Jose Carreras Leukämie-Stiftung, Deutsche Forschungsgemeinschaft (DFG) with specific grant references, LOEWE Hessian Funding Program, Hessen State Ministry for Higher Education, Research and the Arts, and the Cardio-Pulmonary Institute under Germany’s Excellence Strategy. The abstract does not specify how funding supported specific aspects of the work.
The abstract and metadata summarize the approach and major conclusions but omit several methodological specifics: sample sizes, donor characteristics, experimental replication, sequencing depth, and exact analytical pipelines are not reported in the abstract. Any interpretation of findings should therefore consider that full methodological and quantitative detail will be necessary to evaluate reproducibility and generalizability.
This study establishes single-cell measurement of tRNA expression and splicing as a feasible strategy to add a new layer of molecular resolution to studies of human hematopoiesis. By providing an atlas that links tRNA profiles to stemness, differentiation, and lineage identity, the work positions tRNA expression as a biologically relevant component of cellular heterogeneity during blood cell development. The resource and disclosures are made available by the authors; users should consult the full manuscript and deposited materials for complete methods and data access instructions.