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Regensburg 2025 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 25: Cell Mechanics II

BP 25.12: Vortrag

Donnerstag, 20. März 2025, 12:45–13:00, H44

Deformability cytometry for large-scale mechano-genomic screening in interphase and mitosis — •Laura Strampe1, Katarzyna Plak2,3, Christine Schweitzer1, Cornelia Liebers1,2, Buzz Baum3, Jona Kayser1, and Jochen Guck1,21Max-Planck-Zentrum für Physik und Medizin, Erlangen, Germany — 2Biotechnology Center of TU Dresden, Dresden, Germany — 3MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

We demonstrate the scalability of real-time fluorescence and deformability cytometry (RT-FDC) for large-scale cell cycle-resolved mechano-genomic screening. Using RNA interference, we screened 215 kinase and phosphatase genes on their effects on cell mechanics in interphase and mitosis. RT-FDC combines high throughput (up to 100 cells per second) with fluorescence-based cell cycle classification, enabling single-cell mechanical phenotyping of entire populations. We show that cell cycle resolution is essential for identifying genetic regulators of cell mechanics, as stiffness differences between interphase and mitotic cells can obscure genuine knockdown effects or generate false-positive hits. Genes regulating mitotic mechanics or softening cells upon knockdown are particularly likely to be masked. Of the 81 genes identified as affecting cell stiffness, 22 were detected only through cell cycle resolution. These include PRL-1, a cancer metastasis marker with opposing effects across the cell cycle: stiffening interphase cells and softening mitotic cells. This suggests that PRL-1 overexpression in metastatic cells expands the range of mechanical phenotypes during cell cycle progression, facilitating tumor adaptability.

Keywords: Cell mechanics; Screening; Gene regulators; Deformability Cytometry; Mitosis

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