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• W4384388145 abstract "System-level understanding of proteome organization and function requires methods for direct visualization and manipulation of proteins at scale. We developed an approach enabled by high-throughput gene tagging for the generation and analysis of complex cell pools with endogenously tagged proteins. Proteins are tagged with HaloTag to enable visualization or direct perturbation. Fluorescent labeling followed by in situ sequencing and deep learning-based image analysis identifies the localization pattern of each tag, providing a bird's-eye-view of cellular organization. Next, we use a hydrophobic HaloTag ligand to misfold tagged proteins, inducing spatially restricted proteotoxic stress that is read out by single cell RNA sequencing. By integrating optical and perturbation data, we map compartment-specific responses to protein misfolding, revealing inter-compartment organization and direct crosstalk, and assigning proteostasis functions to uncharacterized genes. Altogether, we present a powerful and efficient method for large-scale studies of proteome dynamics, function, and homeostasis." @default.
• W4384388145 created "2023-07-15" @default.
• W4384388145 creator A5030100647 @default.
• W4384388145 creator A5052979776 @default.
• W4384388145 creator A5070112401 @default.
• W4384388145 creator A5074055600 @default.
• W4384388145 creator A5083408012 @default.
• W4384388145 date "2023-07-14" @default.
• W4384388145 modified "2023-10-17" @default.
• W4384388145 title "Pooled tagging and hydrophobic targeting of endogenous proteins for unbiased mapping of unfolded protein responses" @default.
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