<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding"^^ . "All blood and immune cells share the same developmental origin but have diverse functions. Hematopoietic stem cells play a central role in hematopoiesis, as reflected by their ability of multilineage reconstitution of lethally irradiated mice. Recent advancements in DNA barcoding showed that hematopoietic stem cells are not functionally uniform but have heterogenous fates (multilineage, myeloid-erythroid restricted, or differentiation-inactive). The time of hematopoietic stem cells fate specification (embryonically or postnatally) and the stability of distinct fates (at steady state or during transplantation) remain largely unknown. Of note, while DNA barcoding can reveal physiological fates in vivo, it cannot provide molecular insights into fate regulation. To approach these key questions, a next generation barcoding system, PolyloxExpress, was developed. By transcribing the DNA barcodes into RNA, PolyloxExpress allows high-resolution barcoding at both DNA and RNA levels. Applying the PolyloxExpress system in mice and focusing on DNA barcodes, I analyzed hematopoietic stem cell fates at embryonic stages, and found the same fate patterns as in adult stem cells, indicating very early fate determination. To temporally resolve hematopoietic stem cell output in adult life, peripheral blood was repeatedly collected from individual barcoded mice. Distinct stem cell fates remained largely stable over time in both native hematopoiesis and after transplantation, suggesting that hematopoietic stem cell fates are determined cell-intrinsically. To link fates to transcriptomes, I combined PolyloxExpress with single-cell RNA sequencing and established a customized protocol for simultaneous recovery of full-length RNA barcodes and transcriptome. These experiments uncovered that fate-defined hematopoietic stem cell occupied different regions on the transcriptional landscape and have hence, at least partially, distinct transcriptome signatures. These fate-associated signatures were not apparent when analyzing transcriptome alone, which underscores the importance of obtaining fate and transcriptome information together. These data also contain transcripts with uncharacterized functions in stem cells, which may provide a unique resource for future studies. Collectively, PolyloxExpress barcoding revealed coherent hematopoietic stem cell fates which are cell-intrinsically determined and arise around the time of hematopoietic stem cell emergence in the embryo. Fate-associated transcriptome signatures could be identified. Combining PolyloxExpress with single-cell multiomics techniques will provide further insights into molecular aspects of cell fate determination."^^ . "2024" . . . . . . . "Fuwei"^^ . "Shang"^^ . "Fuwei Shang"^^ . . . . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (PDF)"^^ . . . "Fuwei Shang PhD Thesis-Final.pdf"^^ . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (Other)"^^ . . . . . . "indexcodes.txt"^^ . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (Other)"^^ . . . . . . "preview.jpg"^^ . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (Other)"^^ . . . . . . "medium.jpg"^^ . . . "Stem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding (Other)"^^ . . . . . . "small.jpg"^^ . . "HTML Summary of #34609 \n\nStem cell heterogeneity and clonal dynamics explored by PolyloxExpress RNA barcoding\n\n" . "text/html" . . . "570 Biowissenschaften, Biologie"@de . "570 Life sciences"@en . . . "610 Medizin"@de . "610 Medical sciences Medicine"@en . .