Cell organization and disturbance-mapping using image activated cell-profiling (CODIAC)
Krause, M. E. S.; Lou, D.; Li, S.; Jung, H.; Kirrmaier, D.; Gubicza, K.; Schraivogel, D.; Knop, M.; Steinmetz, L. M.
Show abstract
Protein localization and abundance are tightly regulated to maintain cellular homeostasis, and their dysregulation is a hallmark of disease. However, methods that monitor the spatial localization changes of many proteins at once, and at scale, remain limited. Here we introduce CODIAC (Cell Organization and Disturbance-mapping using Image-Activated Cell-profiling), which couples image-enabled cell sorting (ICS) with improved Cas12a-assisted endogenous PCR tagging and machine-learning-guided gating to profile pools of fluorescently tagged proteins. Rather than isolating discrete cell populations, CODIAC reads the "phenotypic fingerprint" of each fluorescently tagged protein, defined by its distribution across a fixed set of image-defined sort bins. The method resolves localization-associated phenotypes, reproducibly profiles complex pools, generalizes to proteins absent from the training set, and detects shifts in protein localization and abundance following chemical perturbation. CODIAC thus extends ICS from outlier screening toward systematic profiling of the spatial proteome and its remodeling under perturbation.
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