[3S00e] 01. ASBA - Part 9

5:05 PM - 5:20 PM JST(8:05 AM - 8:20 AM UTC)

[3ASBA-15-2] Scalable Engineering Tools to Map Protein and Gene Function in Mammalian Cell systems

〇Joonwon Kim (Seoul National University, Seoul, Republic of Korea)

A central challenge in mammalian synthetic biology is to develop scalable engineering tools for interrogating endogenous protein localization, dynamics, and regulation across cellular contexts. Protein localization and regulation are fundamental to understanding cell behavior, yet remain difficult to infer from sequence alone or measure systematically with conventional target-specific antibody-dependent assays. High-throughput Integration of Tags across The Genome (HITAG) addresses this limitation by using Cas9/NHEJ-mediated knock-in to introduce a shared C-terminal epitope tag across many endogenous loci in parallel. This common tagging strategy enables standardized protein-level characterization across distinct biological settings, allowing endogenous proteins to be assayed for basal localization, stress-induced relocalization, and recruitment to stress granules without requiring protein-specific antibodies. By converting many endogenous proteins into a common measurement format, HITAG provides a scalable framework for linking genomic identity to protein localization and context-dependent regulation in mammalian cells. Together, these examples suggest that HITAG can be extended from systematic characterization of endogenous protein behavior to functional protein engineering, using the same native-locus tagging strategy to install regulatory tags and examine their effects in the endogenous cellular context.