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Construction of Cyclic Cell-Penetrating Peptides for Enhanced Penetration of Biological Barriers
Published on: September 19, 2022
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Targeted Subcellular Protein Delivery Using Cleavable Cyclic Cell-Penetrating Peptide-Conjugates
Luise Franz1,2, Anselm F L Schneider1,2, Christian P R Hackenberger3,4
1Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|August 13, 2021
Summary
Cyclic cell-penetrating peptides (cCPPs) enable efficient protein uptake into living cells. This study demonstrates targeted delivery of fluorescent mCherry protein to the nucleus or actin cytoskeleton using cCPP conjugation.
Area of Science:
- Cell biology
- Biochemistry
- Molecular imaging
Background:
- Cell-penetrating peptides facilitate cellular uptake of cargo.
- Cleavable conjugation allows for controlled release and bioavailability of proteins.
- Targeting specific subcellular compartments remains a challenge in protein delivery.
Purpose of the Study:
- To demonstrate the targeted subcellular delivery of mCherry protein using cyclic cell-penetrating peptides (cCPPs).
- To investigate the localization of cCPP-conjugated mCherry in the nucleus and actin cytoskeleton.
- To validate the use of cleavable disulfide bonds for protein release after cellular uptake.
Main Methods:
- Genetic fusion of targeting peptides to mCherry protein.
- Conjugation of targeting peptide-mCherry fusion to cCPP via a cleavable disulfide bond.
- Live cell confocal fluorescence microscopy to observe mCherry localization.
Main Results:
- cCPP-mediated uptake of mCherry into living cells was confirmed.
- Targeting peptides successfully directed mCherry to the nucleus or actin cytoskeleton.
- Cleavable conjugation allowed for protein localization within the cell.
Conclusions:
- cCPP-mediated delivery enables precise subcellular targeting of proteins.
- This approach offers a versatile platform for intracellular protein delivery and imaging.
- The method allows for visualization of protein localization in specific cellular compartments.
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