Direct and Rapid Cytosolic Delivery Using Cell-Penetrating Peptides Mediated by Pyrenebutyrate

2006 ◽  
Vol 1 (5) ◽  
pp. 299-303 ◽  
Author(s):  
Toshihide Takeuchi ◽  
Michie Kosuge ◽  
Akiko Tadokoro ◽  
Yukio Sugiura ◽  
Mayumi Nishi ◽  
...  
Keyword(s):  
Cell Penetrating Peptides ◽  
Cytosolic Delivery ◽  
Cell Penetrating

scholarly journals β-Lactamase Tools for Establishing Cell Internalization and Cytosolic Delivery of Cell Penetrating Peptides

Biomolecules ◽  
2018 ◽  
Vol 8 (3) ◽  
pp. 51 ◽  
Author(s):  
Shane Stone ◽  
Tatjana Heinrich ◽  
Suzy Juraja ◽  
Jiulia Satiaputra ◽  
Clinton Hall ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Cell Penetrating Peptides ◽  
Stable Cell Line ◽  
Intracellular Space ◽  
Biologically Relevant ◽  
Cell Internalization ◽  
Cytosolic Delivery ◽  
Cell Penetrating ◽  
Split Protein

The ability of cell penetrating peptides (CPPs) to deliver biologically relevant cargos into cells is becoming more important as targets in the intracellular space continue to be explored. We have developed two assays based on CPP-dependent, intracellular delivery of TEM-1 β-lactamase enzyme, a functional biological molecule comparable in size to many protein therapeutics. The first assay focuses on the delivery of full-length β-lactamase to evaluate the internalization potential of a CPP sequence. The second assay uses a split-protein system where one component of β-lactamase is constitutively expressed in the cytoplasm of a stable cell line and the other component is delivered by a CPP. The delivery of a split β-lactamase component evaluates the cytosolic delivery capacity of a CPP. We demonstrate that these assays are rapid, flexible and have potential for use with any cell type and CPP sequence. Both assays are validated using canonical and novel CPPs, with limits of detection from <500 nM to 1 µM. Together, the β-lactamase assays provide compatible tools for functional characterization of CPP activity and the delivery of biological cargos into cells.

scholarly journals The Telomerase-Derived Anticancer Peptide Vaccine GV1001 as an Extracellular Heat Shock Protein-Mediated Cell-Penetrating Peptide

2016 ◽  
Author(s):  
Hong Kim ◽  
Eun-Hye Seo ◽  
Seung-Hyun Lee ◽  
Bum-Joon Kim
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Mhc Class Ii ◽  
Biological Effects ◽  
Peptide Vaccine ◽  
Heat Shock Protein 90 ◽  
Cell Penetrating Peptides ◽  
Small Peptides ◽  
Cytosolic Delivery ◽  
Cell Penetrating

Cell-penetrating peptides (CPPs), a group of small peptides capable of promoting the transport of molecular cargo across the plasma membrane, have become important tools in promoting the cellular uptake of exogenously delivered macromolecules. GV1001, a peptide derived from a reverse-transcriptase subunit of telomerase (hTERT) and developed as a vaccine against various cancers, reportedly has unexpected CPP properties. Unlike typical CPPs, such as the HIV-1 TAT peptide, GV1001 enabled the cytosolic delivery of macromolecules such as proteins, DNA and siRNA via extracellular heat shock protein 90 (eHSP90) and 70 (eHSP70) complexes. The eHSP-GV1001 interaction may have biological effects in addition to its cytosolic delivery function. GV1001 was originally designed as a MHC class II-binding cancer epitope, but its CPP properties may contribute to its strong anti-cancer immune response relative to other telomerase peptide-based vaccines. Cell signaling via eHSP-GV1001 binding may lead to unexpected biological effects, such as direct anticancer or antiviral effects. In this review, we focus on the CPP effects of GV1001 bound to eHSP90 and ehsp70.

scholarly journals Unravelling cytosolic delivery of cell penetrating peptides with a quantitative endosomal escape assay

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Serena L. Y. Teo ◽  
Joshua J. Rennick ◽  
Daniel Yuen ◽  
Hareth Al-Wassiti ◽  
Angus P. R. Johnston ◽  
...  
Keyword(s):  
Cell Penetrating Peptides ◽  
Endosomal Escape ◽  
Current Understanding ◽  
Specific Cell ◽  
Intracellular Drug Delivery ◽  
Cytosolic Delivery ◽  
Efficient Delivery ◽  
Cell Penetrating ◽  
Model Protein ◽  
Low Sensitivity

AbstractCytosolic transport is an essential requirement but a major obstacle to efficient delivery of therapeutic peptides, proteins and nucleic acids. Current understanding of cytosolic delivery mechanisms remains limited due to a significant number of conflicting reports, which are compounded by low sensitivity and indirect assays. To resolve this, we develop a highly sensitive Split Luciferase Endosomal Escape Quantification (SLEEQ) assay to probe mechanisms of cytosolic delivery. We apply SLEEQ to evaluate the cytosolic delivery of a range of widely studied cell-penetrating peptides (CPPs) fused to a model protein. We demonstrate that positively charged CPPs enhance cytosolic delivery as a result of increased non-specific cell membrane association, rather than increased endosomal escape efficiency. These findings transform our current understanding of how CPPs increase cytosolic delivery. SLEEQ is a powerful tool that addresses fundamental questions in intracellular drug delivery and will significantly improve the way materials are engineered to increase therapeutic delivery to the cytosol.

scholarly journals Nuclear Localization of Cell-Penetrating Peptides Is Dependent on Endocytosis Rather Than Cytosolic Delivery in CHO Cells

2009 ◽  
Vol 6 (2) ◽  
pp. 337-344 ◽  
Author(s):  
Jennica L. Zaro ◽  
Jacqueline E. Vekich ◽  
Thuy Tran ◽  
Wei-Chiang Shen
Keyword(s):  
Nuclear Localization ◽  
Cho Cells ◽  
Cell Penetrating Peptides ◽  
Cytosolic Delivery ◽  
Cell Penetrating

scholarly journals Cytosolic Delivery of Macromolecules in Live Human Cells Using the Combined Endosomal Escape Activities of a Small Molecule and Cell Penetrating Peptides

2019 ◽  
Vol 14 (12) ◽  
pp. 2641-2651 ◽  
Author(s):  
Jason Allen ◽  
Kristina Najjar ◽  
Alfredo Erazo-Oliveras ◽  
Helena M. Kondow-McConaghy ◽  
Dakota J. Brock ◽  
...  
Keyword(s):  
Small Molecule ◽  
Cell Penetrating Peptides ◽  
Human Cells ◽  
Endosomal Escape ◽  
Cytosolic Delivery ◽  
Cell Penetrating

scholarly journals Monitoring the cytosolic entry of cell-penetrating peptides using a pH-sensitive fluorophore

2015 ◽  
Vol 51 (11) ◽  
pp. 2162-2165 ◽  
Author(s):  
Ziqing Qian ◽  
Patrick G. Dougherty ◽  
Dehua Pei
Keyword(s):  
Cell Penetrating Peptides ◽  
Ph Sensitive ◽  
Cytosolic Delivery ◽  
Cell Penetrating

A simple and effective method for monitoring the cytosolic delivery of cell-penetrating peptides.

scholarly journals Automated high-content imaging for cellular uptake, from the Schmuck cation to the latest cyclic oligochalcogenides

2020 ◽  
Vol 16 ◽  
pp. 2007-2016 ◽  
Author(s):  
Rémi Martinent ◽  
Javier López-Andarias ◽  
Dimitri Moreau ◽  
Yangyang Cheng ◽  
Naomi Sakai ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Cell Penetrating Peptides ◽  
Cellular Systems ◽  
Quantitative Detection ◽  
Response Curves ◽  
Cytosolic Delivery ◽  
Automated Imaging ◽  
Cell Penetrating ◽  
Short Time ◽  
Multiwell Plates

Recent progress with chemistry tools to deliver into living cells has seen a shift of attention from counterion-mediated uptake of cell-penetrating peptides (CPPs) and their mimics, particularly the Schmuck cation, toward thiol-mediated uptake with cell-penetrating poly(disulfide)s (CPDs) and cyclic oligochalcogenides (COCs), here exemplified by asparagusic acid. A persistent challenge in this evolution is the simultaneous and quantitative detection of cytosolic delivery and cytotoxicity in a high-throughput format. Here, we show that the combination of the HaloTag-based chloroalkane penetration assay (CAPA) with automated high-content (HC) microscopy can satisfy this need. The automated imaging of thousands of cells per condition in multiwell plates allows us to obtain quantitative data on not only the fluorescence intensity but also on the localization in a very short time. Quantitative and statistically relevant results can be obtained from dose–response curves of the targeted delivery to selected cells and the cytotoxicity in the same experiment, even with poorly optimized cellular systems.

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