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Tagging and Fusion Proteins

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Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: Sep 13, 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

Programmable protein ligation on cell surfaces.

Christian Kofoed¹, Girum Erkalo¹, Nicholas E S Tay¹

¹Department of Chemistry, Princeton University, Princeton, NJ, USA.

|July 31, 2025

Summary

This summary is machine-generated.

Scientists developed a new molecular device that uses cell surface proteins to make decisions. This programmable system precisely targets cells, enabling localized protein generation for therapeutic or diagnostic applications.

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Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Cell surface landscapes vary by cell type and are altered in disease.
Current strategies often target single surface antigens, limiting specificity.
There is a need for programmable systems that integrate multiple cell-surface features.

Purpose of the Study:

To develop an autonomous decision-making device for targeted protein generation on cell surfaces.
To create a programmable molecular system that integrates multiple cell-surface features for user-defined outputs.

Main Methods:

Utilized proximity-gated protein trans-splicing to generate active proteins from inactive fragments.
Developed a protein-actuator platform for convergent protein ligation on cell surfaces.
Demonstrated modularity and tunability for various inputs, targeting modalities, and outputs.

Main Results:

Achieved highly selective generation of active proteins on designated cell surfaces.
Showcased that generated proteins can remain cell-associated or be released.
Confirmed compatibility with diverse inputs, targeting strategies, and functional outputs.

Conclusions:

The developed protein-actuator platform enables localized interrogation and manipulation of cellular systems.
This programmable molecular system offers a versatile tool for cell-based diagnostics and therapeutics.
The technology allows for precise control over protein generation based on cell-surface recognition.