Aug 2026· Stem Cell Research· Vol 95, pp.
104072
· 0 citations· 5 references
Medicine
TL;DR
A human induced Pluripotent Stem Cell line harboring a doxycycline-inducible Cas9 and an NKX2-1-EGFP-puro reporter via CRISPR/Cas9-mediated homology-directed repair is generated via CRISPR/Cas9-mediated homology-directed repair.
Abstract
NK2 homeobox 1 (NKX2-1), a master regulator robustly expressed in lung, thyroid, and forebrain, is indispensable for specifying lung epithelial fate and serves as a definitive marker of lung progenitors. Here, we generated a human induced Pluripotent Stem Cell (iPSC) line harboring a doxycycline (dox)-inducible Cas9 and an NKX2-1-EGFP-puro reporter via CRISPR/Cas9-mediated homology-directed repair. This dual-function line combines inducible genome editing with real-time tracing of early lung progenitors, enabling their prospective isolation and screening for stage-specific maturation regulators. Therefore, this engineered iCas9-NKX2-1 EGFP line is a key resource for dissecting human lung development, modeling pulmonary disease, and advancing regenerative therapies.
The CRISPR/Cas9 system is utilized to generate a homozygous Mt4 knockout (Mt4-/-) mouse embryonic stem cell (mESC) line that maintains normal morphology, pluripotency, and the ability to differentiate into all three germ layers.
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Key steps include neuro-induction and subsequent maturation using specific growth factors, as well as potential approaches to improve cellular adhesion during maturation, providing a valuable tool for neuroscience research and cellular modeling.
T. Rabin, Anthony Flamier· Journal of Visualized Experi...· 0 citations
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Key applications of iPSC technology in hemato-oncology are summarized, its major advantages and current limitations are discussed, and emerging directions are highlighted, including scalable iPSC-derived blood cell therapies for inherited and acquired bone marrow failure syndromes and leukemia.
Ivan Tesakov, M. Nasri, M. Klimiankou et al.· Frontiers in Immunology· 0 citations
This study demonstrates the integration of CRISPRa with single-cell technologies as a powerful platform for investigating and enriching iPSC-derived NCs, supporting future regenerative strategies across various fields.
Xiao-Le Tong, Marieke Visscher, F. Riemers et al.· Stem Cell Reports· 0 citations
This study has established a highly efficient in vitro model for totipotency induction and highlighted molecular barriers shaping cell fate decisions, providing a platform to dissect the mechanisms governing the pluripotency-to-totipotency transition.
Siling Hu, Ping-Hui Zhu, He Liu et al.· bioRxiv· 0 citations
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