Generation of CD43-P2A-mNeonGreen reporter human pluripotent stem cell line using CRISPR Cas9

This study presents a human induced pluripotent stem cell line engineered to report endogenous CD43 expression by mNeonGreen presence, e.g. enabling visualization of hematopoietic emergence during the endothelial-to-hematopoietic transition. To create this resource, Cas9 ribonucleoprotein targeting the SPN locus and a PCR amplified homology-directed-repair template were used to introduce the reporter sequence, followed by clonal isolation and molecular characterization. The resulting line maintains pluripotency, genomic integrity, and trilineage differentiation potential. This resource accurately reports CD43 + cells during hematopoietic organoid differentiation, supporting applications in studying CD43 + expression dynamics.