Generation of AAVS1 and CLYBL STRAIGHT-IN v2 acceptor human iPSC lines for integrating DNA payloads

Albert Blanch-Asensio; Babet van der Vaart; Mariana Vinagre; Eline Groen; Christiaan Arendzen; Christian Freund; Niels Geijsen; Christine L Mummery; Richard P Davis

doi:10.1016/j.scr.2022.102991

Generation of AAVS1 and CLYBL STRAIGHT-IN v2 acceptor human iPSC lines for integrating DNA payloads

Stem Cell Res. 2023 Feb:66:102991. doi: 10.1016/j.scr.2022.102991. Epub 2022 Dec 5.

Authors

Albert Blanch-Asensio¹, Babet van der Vaart², Mariana Vinagre¹, Eline Groen³, Christiaan Arendzen², Christian Freund², Niels Geijsen¹, Christine L Mummery⁴, Richard P Davis⁵

Affiliations

¹ 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, 2300RC Leiden, the Netherlands.
² Department of Anatomy and Embryology, Leiden University Medical Center, 2300RC Leiden, the Netherlands; LUMC hiPSC Hotel, 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, 2300RC Leiden, the Netherlands.
³ Department of Anatomy and Embryology, Leiden University Medical Center, 2300RC Leiden, the Netherlands.
⁴ 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, 2300RC Leiden, the Netherlands; Department of Applied Stem Cell Technologies, University of Twente, 7500AE Enschede, the Netherlands.
⁵ 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, 2300RC Leiden, the Netherlands. Electronic address: r.p.davis@lumc.nl.

PMID: 36495703
DOI: 10.1016/j.scr.2022.102991

Abstract

STRAIGHT-IN is a platform to precisely integrate DNA payloads into the genome of cells, including hiPSCs. Here, we generated two hiPSC acceptor lines each with one copy of an upgraded landing pad (LP). This improved design allows more efficient (∼100 %) and rapid (∼2-3 weeks) generation of genetically modified hiPSC lines containing the desired payloads. This new LP version was inserted into either the AAVS1 (LUMCi004-A-1) or CLYBL (LUMCi004-A-2) safe harbour loci in the hiPSC line, LUMC0099iCTRL04. The resulting lines can be used for the targeted integration of a wide range of transgenes, thereby making them suitable for numerous research applications.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA / genetics
Humans
Induced Pluripotent Stem Cells*
Transgenes

Substances

DNA
CLYBL protein, human