Versatile Labeling and Detection of Endogenous Proteins Using Tag-Assisted Split Enzyme Complementation

• DOI: 10.1021/acschembio.0c00925
• Publication Date: 2021-03-18T18:03:24Z
• Authors: Suraj Makhija; David Brown; Rachel M. Rudlaff; Julia K. Doh; Struan Bourke; Yina Wang; Shuqin Zhou; Rasmi Cheloor-Kov...; Bo Huang
• Keywords: Protein-fragment complementation assay; Protein tag

Recent advances in genome engineering have expanded our capabilities to study proteins their natural states. In particular, the ease and scalability of knocking-in small peptide tags has enabled high throughput tagging analysis endogenous proteins. To improve enrichment capacities expand functionality knock-ins using short tags, we developed tag-assisted split enzyme complementation (TASEC) approach, which uses two orthogonal cognate binders conditionally drive a upon labeled protein expression. Using this engineered optimized HaloTag system (TA-splitHalo) demonstrated its versatile applications improving efficiency knock-in cell enrichment, detection protein-protein interaction, isolation biallelic gene edited cells through multiplexing.