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Optogenetic Blue GENEs engineered into a human safe harbor locus

Title: Optogenetic Blue GENEs engineered into a human safe harbor locus
Authors: Geidies, Alexander; Nieke, Marius; Witte, Nils; McLean, Benjamin J; Evangelopoulou, Maria; Koh, Cha San; Kuschel, Stefanie; Stölting, Frauke; Zurbriggen, Matias D; Beyer, Hannes M
Contributors: Volkswagen Foundation; Dr Rolf M. Schwiete Foundation; Research Executive Agency; Deutsche Forschungsgemeinschaft; Germany ́s Excellence Strategy
Source: Nucleic Acids Research ; volume 54, issue 2 ; ISSN 0305-1048 1362-4962
Publisher Information: Oxford University Press (OUP)
Publication Year: 2026
Description: Crafting synthetic in vitro tissues with mammalian cells faces a shortage of methods to define spatial features. Optogenetic tissue engineering can provide the desired spatial and temporal control but requires stable genomic engineering to support long-term cultivation and high response resolution. Here, we developed BlueGENEs, a set of optimized optogenetic gene switches. BlueGENEs support rapid, stable cell line generation, including precision engineering into the human AAVS1 safe harbor locus. By combining a designer endonuclease and a phage integrase, the approach overcomes gene-disruptive effects of random gene delivery and enables reproducible cell line development. BlueGENEs comprise an optogenetic blue light-responsive gene switch, a synthetic response promoter, and selection strategies serving broad use scenarios. We generated various human cell lines for optical control of apoptotic cell fate, 3D tissue formation, and signals promoting cytoskeletal remodeling. Our results demonstrate the integration of optogenetic cells with bioprinting technologies, illustrating the potential of BlueGENEs in advancing the synthesis of de novo or patient-derived in vitro model systems.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1093/nar/gkaf1461
Availability: https://doi.org/10.1093/nar/gkaf1461; https://academic.oup.com/nar/article-pdf/54/2/gkaf1461/66411263/gkaf1461.pdf
Rights: https://creativecommons.org/licenses/by-nc/4.0/
Accession Number: edsbas.FC485FF9
Database: BASE