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The role of aphidicolin-sensitive fragile sites in iPSCs genetic instability
https://doi.org/10.25557/2073-7998.2025.08.94-97
Abstract
Introduction. One of the causes of genetic instability in iPSCs is replication stress (RS). RS leads to the formation of chromatid breaks and gaps in regions of constitutive fragile sites (cFS), which are «hotspots» for chromosomal rearrangements. The process of mitotic DNA synthesis (MiDAS) serves to restore genome integrity under RS and acts as a marker of cFS.
Objective: mapping chromosomal breaks and MiDAS sites in iPSCs.
Methods. RS was induced using aphidicolin and caffeine. MiDAS regions were detected by EdU incorporation at early mitotic stages. Differential staining was performed using DAPI and actinomycin D.
Results. Three iPSC lines (P1L5, P12L3, P16L1) were analyzed. Among 800 mapped chromosomal breaks, 33 cFS were identified. Both previously described and iPSC-specific cFS were characterized. cFS co-localized with breakpoints of recurrent karyotypic abnormalities in iPSCs were identified.
Conclusions. For the first time, a repertoire of cFS in iPSCs and their associations with recurrent rearrangements has been described.
Keywords
Supporting agencies: The work was carried out within the framework of the state assignment of the Ministry of Science and Higher Education of Russia for the Research Centre for Medical Genetics
About the Authors
V. O. Pozhitnova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
Victoria O. Pozhitnova
115522; 1, Moskvorechie st.; Moscow
A. V. Kislova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
P. S. Sviridov
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
V. V. Sviridova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
L. I. Gumerova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
D. S. Kiselev
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
D. G. Zheglo
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
E. S. Voronina
Research Centre for Medical Genetics
Russian Federation
Russian Federation
115522; 1, Moskvorechie st.; Moscow
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Review
For citations:
Pozhitnova V.O., Kislova A.V., Sviridov P.S., Sviridova V.V., Gumerova L.I., Kiselev D.S., Zheglo D.G., Voronina E.S. The role of aphidicolin-sensitive fragile sites in iPSCs genetic instability. Medical Genetics. 2025;24(8):94-97. (In Russ.) https://doi.org/10.25557/2073-7998.2025.08.94-97
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