Systemic nucleic acid delivery to hematopoietic stem and progenitor cells (HSPCs) remains a major barrier to in vivo gene therapy due to hepatic sequestration of lipid nanoparticles (LNPs). Here, we report a scalable, ligand-free HSPC-tropic LNP enabling efficient mRNA delivery to human HSPCs in vivo without conditioning. In humanized NBSGW mice, the LNP preferentially delivered mRNA to human HSPCs and early myeloid progenitors, with limited delivery to lymphoid and non-hematopoietic cells. Single-cell transcriptomic and multiomic analyses revealed no broad transcriptional perturbation associated with delivery. Compared with the SM-102 LNP, the HSPC-tropic LNP achieved greater mRNA delivery to human HSPCs, while exhibiting substantially reduced hepatic delivery. The LNP also enabled functional delivery of Cas9 mRNA and β2M-targeting sgRNA to human CD34+ HSPCs, resulting in efficient gene editing and reduced β2M expression. These findings establish a ligand-free platform for functional RNA delivery and genome engineering in human HSPCs.
