2. Academic Validation
  3. Protocol for quantifying PARP inhibitor-induced changes in PARP1 dynamics and activity through live-cell imaging

Protocol for quantifying PARP inhibitor-induced changes in PARP1 dynamics and activity through live-cell imaging

  • STAR Protoc. 2025 Sep 25;6(4):104106. doi: 10.1016/j.xpro.2025.104106.
Petar-Bogomil Kanev 1,  Sylvia Varhoshkova 2,  Aleksander Aleksandrov 3,  Stoyno Stoynov 2,  Radoslav Aleksandrov 4
Affiliations

Affiliations

  • 1 Laboratory of Genomic Stability, Institute of Molecular Biology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str. Bl.21, 1113 Sofia, Bulgaria. Electronic address: pkanev@bio21.bas.bg.
  • 2 Laboratory of Genomic Stability, Institute of Molecular Biology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str. Bl.21, 1113 Sofia, Bulgaria.
  • 3 Department of Gene Regulation, Institute of Molecular Biology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str. Bl.21, 1113 Sofia, Bulgaria.
  • 4 Laboratory of Genomic Stability, Institute of Molecular Biology, Bulgarian Academy of Sciences, Acad. G. Bonchev Str. Bl.21, 1113 Sofia, Bulgaria. Electronic address: raleksandrov@bio21.bas.bg.
PMID: 41014565 DOI: 10.1016/j.xpro.2025.104106
Abstract

Poly(ADP-ribose) polymerase 1 (PARP1) is a crucial DNA repair protein and a target of PARP inhibitors (PARPi), which suppress its catalytic activity while also altering its association with damaged chromatin. Here, we present a live-cell imaging protocol for obtaining high-quality kinetics of fluorescently labeled PARP1 and poly(ADP-ribose) (PAR)-binding proteins at micro-irradiation-induced DNA damage sites in untreated and PARPi-treated cells. This approach can be easily adapted to Other DNA repair proteins to gain insight into the mechanism of action of diverse DNA repair-targeting drugs. For complete details on the use and execution of this protocol, please refer to Kanev et al.1.

Keywords

Biophysics; Cancer; Cell Biology; Cell culture; Cell-based Assays; Microscopy; Molecular Biology.

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