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

ACS Chem Biol. 2021 Apr 16;16(4):671-681. doi: 10.1021/acschembio.0c00925. Epub 2021 Mar 18.

Abstract

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

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Enzymes / metabolism*
  • Flow Cytometry
  • Fluorescent Dyes / chemistry
  • HEK293 Cells
  • Humans
  • Protein Binding
  • Proteins / metabolism*

Substances

  • Enzymes
  • Fluorescent Dyes
  • Proteins