CRISPR-Cas12a-integrated transgenes in genomic safe harbors retain high expression in human hematopoietic iPSC-derived lineages and primary cells

We identified and characterized potential integration safe harbor sites (SHS) in human cells. Using the CRISPR-MAD7 system, we integrated transgenes at these genomic sites in iPSC, primary T and NK cells, and Jurkat cell line, and demonstrated efficient and stable expression at these loci. Subsequently, we validated the differentiation capabilities of engineered iPSC towards CD34+ hematopoietic stem and progenitor cells (HSPC), lymphoid progenitors (LPC), and natural killer (NK) cells, and showed that transgene expression was retained in these lineages.