A novel “chassis” PSC line for precise safe harbor site integration via selection-aided RMCE

We describe here a human pluripotent stem cell line (hPSC) designed to serve as an acceptor (a “chassis” cell line) for the recombination-mediated cassette exchange (RMCE)-mediated precise insertion of DNA fragments into a safe harbor locus. To enable the use of Flp recombinase for directional inser...

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Published inStem cell research Vol. 87; p. 103758
Main Authors Ay, Merve, Veleva, Desi, Chowdhury, Mohammad M., Prowse, Andrew B.J., Ovchinnikov, Dmitry A.
Format Journal Article
LanguageEnglish
Published England Elsevier B.V 01.09.2025
Elsevier
Subjects
Cell Line
Humans
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
Recombination, Genetic
Online AccessGet full text
ISSN1873-5061
1876-7753
1876-7753
DOI10.1016/j.scr.2025.103758

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Summary:We describe here a human pluripotent stem cell line (hPSC) designed to serve as an acceptor (a “chassis” cell line) for the recombination-mediated cassette exchange (RMCE)-mediated precise insertion of DNA fragments into a safe harbor locus. To enable the use of Flp recombinase for directional insertion, we targeted one of the AAVS1 safe harbor alleles with an excisable negative selection cassette flanked with FRT and FRT3 sites. The generated hPSC line displayed typical colony morphology, pluripotency signatures and normal karyotype. We expect it to provide a useful AAVS1 safe harbor “landing site” for FRT/FRT3-flanked transgenes.
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ISSN:1873-5061
1876-7753
1876-7753
DOI:10.1016/j.scr.2025.103758