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The  Reprimo  gene family comprises a group of single-exon genes for which their physiological function remains poorly understood. Heretofore, mammalian  Reprimo  ( RPRM ) has been described as a putative p53-dependent tumor suppressor gene that functions at the G2/M cell cycle checkpoint. Another family member,  Reprimo-like  ( RPRML ), has not yet an established role in physiology or pathology. Importantly,  RPRML  expression pattern is conserved between zebrafish and human species. Here, using CRISPR-Cas9 and antisense morpholino oligonucleotides, we disrupt the expression of  rprml  in zebrafish and demonstrate that its loss leads to impaired definitive hematopoiesis. The formation of hemangioblasts and the primitive wave of hematopoiesis occur normally in absence of  rprml . Later in development there is a significant reduction in erythroid-myeloid precursors (EMP) at the posterior blood island (PBI) and a significant decline of definitive hematopoietic stem/progenitor cells (HSPCs). Furthermore, loss of  rprml  also increases the activity of caspase-3 in endothelial cells within the caudal hematopoietic tissue (CHT), the first perivascular niche where HSPCs reside during zebrafish embryonic development. Herein, we report an essential role for  rprml  during hematovascular development in zebrafish embryos, specifically during the definitive waves of hematopoiesis, indicating for the first time a physiological role for the  rprml  gene.

The Reprimo gene family member, reprimo-like (rprml), is required for blood development in embryonic zebrafish