Lin(−) PU.1(dim)GATA‐1(−) defines haematopoietic stem cells with long‐term multilineage reconstitution activity

Despite extensive characterization of the state and function of haematopoietic stem cells (HSCs), the use of transcription factors to define the HSC population is still limited. We show here that the HSC population in mouse bone marrow can be defined by the distinct expression levels of Spi1 and Gata1. By using a double fluorescence knock‐in mouse model, PGdKI, in which the expression levels of PU.1 and GATA‐1 are indicated by the expression of GFP and mCherry, respectively, we uncover that the HSCs with lymphoid and myeloid repopulating activity are specifically enriched in a Lin(−)PU.1(dim)GATA‐1(−) (LPG) cell subset. In vivo competitive repopulation assays demonstrate that bone marrow cells gated by LPG exhibit haematopoietic reconstitution activity which is comparable to that of classical Lin(−)Sca1(+)c‐kit(+) (LSK). The integrated analysis of single‐cell RNA sequence data from LPG and LSK‐gated cells reveals that a transcriptional network governed by core TFs contributes to regulation of HSC multipotency. These discoveries provide new clues for HSC characterization and functional study.