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Related Experiment Video

Updated: Oct 13, 2025

Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT
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Protein Delivery by PTDs/CPPs.

Caroline Palm-Apergi1, Steven F Dowdy2

  • 1Department of Laboratory Medicine, Biomolecular and Cellular Medicine, Karolinska Institutet, Stockholm, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|November 12, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a method to deliver proteins into cells using TAT-fusion proteins. This technique enables manipulation of cell biology for research and potential therapeutic applications.

Keywords:
Cell-penetrating peptide (CPP)Protein transductionProtein transduction domain (PTD)TAT

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Area of Science:

  • Cell biology
  • Protein transduction
  • Biotechnology

Background:

  • Protein delivery into cells is crucial for manipulating cellular functions in various research and therapeutic settings.
  • TAT peptide transduction domain (PTD), also known as cell-penetrating peptide (CPP), facilitates the entry of proteins into cells.
  • Existing methods for protein delivery have limitations in scope and efficiency.

Purpose of the Study:

  • To describe a method for generating and purifying TAT-fusion proteins for efficient cellular delivery.
  • To highlight the versatility of TAT-fusion proteins for delivering diverse functional proteins.

Main Methods:

  • Generating TAT-fusion proteins by cloning into the pTAT-HA plasmid.
  • Expressing fusion proteins in E. coli.
  • Purifying proteins using 6-His affinity tag and Nickel-NTA chromatography.
  • Final purification via Ion Exchange Fast Protein Liquid Chromatography (IEX FPLC).

Main Results:

  • Successfully generated and purified TAT-fusion proteins.
  • Demonstrated the capability of TAT-fusion proteins to deliver various protein types, including enzymes and transcription factors.
  • Established a robust purification protocol for TAT-fusion proteins.

Conclusions:

  • TAT-fusion proteins offer a powerful tool for protein transduction into cells.
  • This method allows for the manipulation of cell biology in research and preclinical models.
  • The developed technique has potential implications for future therapeutic applications in human diseases.