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

A human induced pluripotent stem cell line was engineered with a CD43-P2A-mNeonGreen reporter that enables accurate visualization of CD43+ cells during hematopoietic organoid differentiation.

• Why it matters: Tracking endogenous CD43 expression is needed to study how hematopoietic cells emerge, particularly during the endothelial-to-hematopoietic transition.
• What they did: Cas9 ribonucleoprotein targeting the SPN locus and a PCR-amplified homology-directed-repair template were used, followed by clonal isolation and molecular characterization.
• The result: The edited line retains pluripotency and differentiation potential and accurately reports CD43+ cells during hematopoietic organoid differentiation, supporting studies of CD43+ expression dynamics.

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