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

Stem Cell Research (Feb 2023)

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

Affiliations

Albert Blanch-Asensio: 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
Babet van der Vaart: 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
Mariana Vinagre: 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
Eline Groen: Department of Anatomy and Embryology, Leiden University Medical Center, 2300RC Leiden, the Netherlands
Christiaan Arendzen: 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
Christian Freund: 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
Niels Geijsen: 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
Christine L. Mummery: 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
Richard P. Davis: 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; Corresponding author.

Journal volume & issue: Vol. 66
p. 102991

Abstract

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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.

Published in Stem Cell Research

ISSN: 1873-5061 (Print); 1876-7753 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: https://www.journals.elsevier.com/stem-cell-research

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