CaMK2rep: A Highly Sensitive Genetically Encoded Biosensor for Monitoring CaMKII Activity in Mammalian Cells.

ABSTRACT: Accurately monitoring calcium/calmodulin-dependent protein kinase II (CaMKII) activity in cells remains a significant challenge due to the limited sensitivity and narrow dynamic range of existing genetically encoded sensors. Here, we introduce CaMK2rep, a novel phosphorylation-based biosensor that enables robust, specific, and high-sensitivity detection of CaMKII activity. CaMK2rep is designed with two tandem CaMKII consensus sites embedded within the native sequence context of synapsin, and its phosphorylation is detected via a phospho-specific antibody, allowing both biochemical and morphological analyses. We validated CaMK2rep in HeLa cells and cultured hippocampal neurons, demonstrating a near-linear response to CaMKII expression levels and to stimulation intensity, and no detectable c

SUBMITTER: Martinez-Blanco E 

PROVIDER: S-EPMC12461680 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature