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Generation of a STRAIGHT-IN Dual AAVS1 hiPSC line with orthogonal landing pads for versatile DNA payload integration
Babet van der Vaart1, Albert Blanch-Asensio2, Niels Geijsen2
1Department of Anatomy and Embryology, Leiden University Medical Center, 2300RC Leiden, the Netherlands; Leiden hiPSC Centre, Department of Anatomy and Embryology, Leiden University Medical Center, 2300RC Leiden, the Netherlands; The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, the Netherlands.
Abstract:
The STRAIGHT-IN platform is designed for facile genomic integration of DNA payloads into human induced pluripotent stem cells (hiPSCs) that contain a pre-inserted landing pad (LP). Here, we expanded the versatility of STRAIGHT-IN by introducing an additional, orthogonal LP into the unmodified allele of the safe harbor locus AAVS1. Specifically, we targeted the hiPSC line LUMC0099iCTRL04_AAVS1-bxb-v2 (hPSCreg LUMCi004-A-1), which already carried one LP. The resulting STRAIGHT-IN AAVS1 Dual line can integrate two independent DNA payloads in parallel, expanding the applicability of the platform for complex genomic engineering applications.
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