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Method Article
Matthias Altmeyer
Altmeyer Lab, Department of Molecular Mechanisms of Disease, University of Zurich, Switzerland
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Current methods employed to evaluate PARP inhibitor (PARPi) toxicity can be time consuming, have limited sensitivity, may not be well suited for screening purposes, or yield results from cell population averages rather than single cell information. To overcome these limitations we provide a detailed protocol to assess PARPi toxicity at multiple phenotypic levels based on a high-content microscopy workflow. This approach takes two days and allows for sensitive cell cycle resolved analyses of PARPi-evoked DNA damage response signaling and PARP trapping. The approach can be applied to evaluate conditions of PARPi hypersensitivity and resistance, and to obtain insights in the cellular mechanisms of drug synergism. Moreover, the described workflow is compatible with high-throughput screening for the discovery and characterization of functional interactions between PARPi and other clinically relevant drugs.
Keywords
PARP inhibitor, DNA damage response, BRCA1, BRCA2, synthetic lethality, PARP inhibitor sensitivity and resistance, ATR inhibitor, SCF inhibitor
Figure 1
The authors declare no competing financial interests.
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Associated Publications
Analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy reveals mechanisms of sensitivity and resistance
by Jone Michelena, Aleksandra Lezaja, Federico Teloni, +3
Nature Communications (13 December, 2018)
Method Article
Matthias Altmeyer
Altmeyer Lab, Department of Molecular Mechanisms of Disease, University of Zurich, Switzerland
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 28 Jan, 2019
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Current methods employed to evaluate PARP inhibitor (PARPi) toxicity can be time consuming, have limited sensitivity, may not be well suited for screening purposes, or yield results from cell population averages rather than single cell information. To overcome these limitations we provide a detailed protocol to assess PARPi toxicity at multiple phenotypic levels based on a high-content microscopy workflow. This approach takes two days and allows for sensitive cell cycle resolved analyses of PARPi-evoked DNA damage response signaling and PARP trapping. The approach can be applied to evaluate conditions of PARPi hypersensitivity and resistance, and to obtain insights in the cellular mechanisms of drug synergism. Moreover, the described workflow is compatible with high-throughput screening for the discovery and characterization of functional interactions between PARPi and other clinically relevant drugs.
Figure 1
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Associated Publications
Analysis of PARP inhibitor toxicity by multidimensional fluorescence microscopy reveals mechanisms of sensitivity and resistance
by Jone Michelena, Aleksandra Lezaja, Federico Teloni, +3
Nature Communications (13 December, 2018)