Generation of a human embryonic stem cell line (SMUDHe010-A-3F) with Cas9 expression cassette integrated at the AAVS1 locus via CRISPR/Cas9-mediated homologous recombination
Summary
Cas9, an RNA-guided nuclease, enables precise genome editing by recognizing sgRNA-complementary sequences and cleaving target DNA. In this study, we used CRISPR/Cas9-mediated homologous recombination to integrate a loxP-flanked STOP cassette-controlled Cas9 expression framework (LSL-Cas9) into the AAVS1 safe-harbor locus of human embryonic stem cells. The resulting cell line, SMUDHe010-A-3F, allows Cre-dependent activation of Cas9 but remains inactive in the absence of Cre recombinase. Karyotype and tri-lineage differentiation confirmed genomic stability and pluripotency. This line provides a valuable platform for organoid gene editing and studies of human development and disease. Copyright © 2025 The Author(s). Published by Elsevier B.V. All rights reserved.
| Authors | Wang Y, Zheng W, Qiu B, Chen Q, Yang T, Zhou S, Liu J, Yang B |
|---|---|
| Journal | Stem cell research |
| Publication Date | 2025 Dec 9;90:103882 |
| PubMed | 41418729 |
| DOI | 10.1016/j.scr.2025.103882 |