Ophir Shalem

Keywords pooled tagging proteostasis in situ sequencing protein misfolding protein localization hydrophobic targeting Introduction Large-scale multiplexed imaging and gene perturbation technologies have transformed mammalian cell biology but are predominantly limited to nucleic acids. This is due to the ease of generating specific targeting reagents via sequence complementarity, which does not extend to the proteins.

Abstract The use of CRISPR-associated enzymes in iPSC-derived neurons for precise gene targeting and high-throughput gene perturbation screens offers great potential but presents unique challenges compared to dividing cell lines.

Abstract The subcellular organization of proteins carries important information on cellular state and gene function, yet currently there are no technologies that enable accurate measurement of subcellular protein localizations at scale. Here we develop an approach for pooled endogenous protein tagging using prime editing, which coupled with an optical readout and sequencing, provides a snapshot of proteome organization in a manner akin to perturbation-based CRISPR screens.