Versatile Labeling and Detection of Endogenous Proteins Using Tag-Assisted Split Enzyme Complementation.
Suraj MakhijaDavid BrownRachel M RudlaffJulia K DohStruan BourkeYina WangShuqin ZhouRasmi Cheloor-KovilakamBo HuangPublished in: ACS chemical biology (2021)
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.
Keyphrases
- protein protein
- high throughput
- single cell
- small molecule
- induced apoptosis
- genome wide
- loop mediated isothermal amplification
- label free
- real time pcr
- copy number
- stem cells
- cell cycle arrest
- dna methylation
- cell therapy
- intellectual disability
- cell death
- gene expression
- mesenchymal stem cells
- pet imaging
- computed tomography
- heat shock
- oxidative stress
- heat stress