MCerulean3-Based Cameleon Sensor to Explore Mitochondrial Ca²⁺ Dynamics In Vivo.

ABSTRACT: Genetically Encoded Ca²⁺ Indicators (GECIs) are extensively used to study organelle Ca²⁺ homeostasis, although some available probes are still plagued by a number of problems, e.g., low fluorescence intensity, partial mistargeting, and pH sensitivity. Furthermore, in the most commonly used mitochondrial Förster Resonance Energy Transfer based-GECIs, the donor protein ECFP is characterized by a double exponential lifetime that complicates the fluorescence lifetime analysis. We have modified the cytosolic and mitochondria-targeted Cameleon GECIs by (1) substituting the donor ECFP with mCerulean3, a brighter and more stable fluorescent protein with a single exponential lifetime; (2) extensively modifying the constructs to improve targeting efficiency and fluorescence cha

SUBMITTER: Greotti E 

PROVIDER: S-EPMC6581653 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature