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Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks
Published on: December 24, 2016
Improved tagging strategy for protein identification in mammalian cells
Agnieszka Bialkowska1, Xian-Yang Zhang, Jakob Reiser
1Gene Therapy Program, Department of Medicine, Louisiana State University Health Sciences Center, New Orleans, Louisiana 70112, USA. abialk@lsuhsc.edu
BMC Genomics
|September 6, 2005
Summary
Researchers developed new genetic tools for tagging full-length endogenous proteins in mammalian cells using enhanced green fluorescent protein (EGFP). This method allows for studying protein localization and interactions without affecting protein function.
Area of Science:
- Molecular Biology
- Cell Biology
Background:
- Mammalian cell protein tagging enables expression of full-length endogenous proteins as fluorescent fusions.
- Existing methods can be improved for greater efficiency and accuracy.
Purpose of the Study:
- To develop and present improved genetic tools for protein tagging in mammalian cells.
- To facilitate the study of endogenous proteins using fluorescent tags.
Main Methods:
- Utilized a mobile genetic element with an artificial exon encoding a protein tag.
- Inserted the artificial exon into introns of cellular genes via lentiviral vectors.
- Introduced enhanced green fluorescent protein (EGFP) tags into expressed genes.
Main Results:
- Achieved expression of bona fide tripartite fusion proteins.
- Demonstrated that the EGFP tag does not alter the subcellular localization of tagged proteins.
- Successfully tagged full-length endogenous proteins in mammalian cells.
Conclusions:
- The developed tools enable protein discovery and investigation of subcellular distribution and function.
- Facilitates studies on protein purification and protein-protein interactions.
- Provides a robust method for endogenous protein analysis in mammalian systems.
