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Related Experiment Video

Updated: Nov 12, 2025

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks

Published on: December 24, 2016

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

Suraj Makhija¹, David Brown², Rachel M Rudlaff²

¹UC Berkeley - UCSF Joint Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, California 94143, United States.

ACS Chemical Biology

|March 18, 2021

Summary

This summary is machine-generated.

We developed a novel tag-assisted split enzyme complementation (TASEC) method for improved analysis of endogenous proteins. This approach enhances knock-in cell enrichment, protein interaction studies, and gene editing applications.

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Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Genome engineering allows studying proteins in natural states.
Small peptide tags facilitate high-throughput endogenous protein analysis.
Current methods have limitations in enrichment capacity and functionality.

Purpose of the Study:

To develop an improved method for endogenous protein analysis using short tags.
To enhance knock-in cell enrichment and expand functional applications.
To introduce the tag-assisted split enzyme complementation (TASEC) approach.

Main Methods:

Developed the tag-assisted split enzyme complementation (TASEC) approach.
Engineered and optimized the tag-assisted split HaloTag complementation system (TA-splitHalo).
Utilized two orthogonal small peptide tags and cognate binders for enzyme complementation.

Main Results:

Demonstrated improved efficiency in knock-in cell enrichment.
Successfully detected protein-protein interactions.
Enabled isolation of biallelic gene edited cells via multiplexing.

Conclusions:

TASEC approach significantly enhances endogenous protein studies.
TA-splitHalo system offers versatile applications in cell enrichment and gene editing.
This method provides a scalable and functional tool for molecular biology research.