bioRxivBioengineeringPreprint
Paralog-specific intrabodies for PSD-93 and SAP102 expand the molecular toolkit to resolve excitatory synapse organization
Christelle Breillat, Ellyn Renou, Manon DarribereiD, Charlotte RimbaultiD, Vincent TalentoniD, Agathe Ecoutin, Sophie Daburon, Christel PoujoliD, Daniel ChoquetiD, Cameron D. MackerethiD, Matthieu SainlosiD
Abstract
A scarcity of live, paralog-specific tools has limited analysis of PSD-MAGUKs at excitatory synapses. To address this gap, we engineered small, 10FN3-derived binders that selectively recognize PSD-93 and SAP102 -alongside an enhanced PSD-95 reagent- and converted them into regulated, gene-encoded intrabodies for endogenous imaging. Through sequence-guided selection and targeted optimization, we obtained high-specificity reagents that label their native targets in neurons with minimal perturbation and support multiplexed live-cell and advanced imaging modalities. This toolkit enables differential visualization of MAGUK paralogs at native levels and provides a practical route to dissect their distinct contributions to synapse organization and plasticity.
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