Novel fluorescence resonance energy transfer‐based reporter reveals differential calcineurin activation in neonatal and adult cardiomyocytes

Novel fluorescence resonance energy transfer‐based genetically encoded reporters of calcineurin are constructed by fusing the two subunits of calcineurin with P2A‐based linkers retaining the expected native conformation of calcineurin. Calcineurin reporters display robust responses to calcium transients in HEK293 cells. The sensor responses are correlated with NFATc1 translocation dynamics in HEK293 cells. The sensors are uniformly distributed in neonatal myocytes and respond efficiently to single electrically evoked calcium transients and show cumulative activation at frequencies of 0.5 and 1 Hz. In adult myocytes, the calcineurin sensors appear to be localized to the cardiac z‐lines, and respond to cumulative calcium transients at frequencies of 0.5 and 1 Hz.

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