Patient-specific induced pluripotent stem cells (iPSCs) capable of differentiation into required cell type are a promising model for studying various pathological processes and development of new therapeutic approaches. However, no conventional strategies for using iPSCs in disease research have been established yet. Genetically encoded biosensors can be used for monitoring messenger molecules, metabolites, and enzyme activity in real time with the following conversion of the registered signals in quantitative data, thus allowing evaluation of the impact of certain molecules on pathology development. We have generated a number of transgenic cell lines, encoding various biosensors’ sequences in the safe-harbor AAVS1 locus.
These sensors allow to measure the following processes:
– Oxidative state of the cell (through measurement of the relative H
2O
2 levels in cytoplasm/mitochondria, or reduced/oxidized glutathione ratio);
– Apoptosis (through caspase-3 activity);
– ER-stress and UPR (XBP1 activity, ATF6 activity)
– Ca
2+-imaging