Engineering STRAIGHT-IN single and dual lines in the male iPS11 parental line for programmable DNA integration.

STRAIGHT-IN is a genome engineering platform that enables precise integration of DNA payloads into mammalian genomes, including hiPSCs. In this study, we generated three hiPSC acceptor lines containing either one (single) or two (dual) landing pads. These landing pads support efficient, seamless integration of DNA cargos with single-copy control and a near-scarless genomic footprint. All landing pads were targeted to the CLYBL genomic safe harbor locus in the male hiPSC line iPS11. The resulting acceptor lines offer a versatile resource for the controlled genomic integration of diverse transgenes, making them broadly applicable to a wide range of applications.