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

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 expa...

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Bibliographic Details
Published in:ACS chemical biology 2021-04, Vol.16 (4), p.671-681
Main Authors: Makhija, Suraj, Brown, David, Rudlaff, Rachel M, Doh, Julia K, Bourke, Struan, Wang, Yina, Zhou, Shuqin, Cheloor-Kovilakam, Rasmi, Huang, Bo
Format: Article
Language:English
Subjects:
Enzymes - metabolism
Flow Cytometry
Fluorescent Dyes - chemistry
HEK293 Cells
Humans
Protein Binding
Proteins - metabolism
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Summary: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.
ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.0c00925