ArticleBiochemistry2013
Improved assays for determining the cytosolic access of peptides, proteins, and their mimetics.
Article in Biochemistry, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Membrane-Traversing Peptides: Mechanistic Landscapes, Assays, and Applications.Chemical reviews · 2026Review
- Protein Modifications for Cellular Protein Delivery.Chemical reviews · 2026Review
- Packaged delivery of CRISPR-Cas9 ribonucleoproteins accelerates genome editing.Nucleic acids research · 2025Article
- Recent advances in methods for quantifying the cell penetration of macromolecules.Current opinion in chemical biology · 2024Review
- Cell-permeable chameleonic peptides: Exploiting conformational dynamics in de novo cyclic peptide design.Current opinion in structural biology · 2023Review
- Importance of two-dimensional cation clusters induced by protein folding in intrinsic intracellular membrane permeability.RSC chemical biology · 2022Article
- Cytosolic Delivery of Argininosuccinate Synthetase Using a Cell-Permeant Miniature Protein.ACS central science · 2021Article
- A critical analysis of methods used to investigate the cellular uptake and subcellular localization of RNA therapeutics.Nucleic acids research · 2020Review
- In Vitro Assays: Friends or Foes of Cell-Penetrating Peptides.International journal of molecular sciences · 2020Review
- Quantitative measurement of cytosolic penetration using the chloroalkane penetration assay.Methods in enzymology · 2020Article
- Understanding Cell Penetration of Cyclic Peptides.Chemical reviews · 2019Review
- Engineered Polymeric Materials for Biological Applications: Overcoming Challenges of the Bio-Nano Interface.Polymers · 2019Review
- Trapped! A Critical Evaluation of Methods for Measuring Total Cellular Uptake versus Cytosolic Localization.Bioconjugate chemistry · 2019Review
- HOPS-dependent endosomal fusion required for efficient cytosolic delivery of therapeutic peptides and small proteins.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Fluorescence Correlation Spectroscopy Reveals Efficient Cytosolic Delivery of Protein Cargo by Cell-Permeant Miniature Proteins.ACS central science · 2018Article
- Cell Penetration Profiling Using the Chloroalkane Penetration Assay.Journal of the American Chemical Society · 2018Article
- Emerging Methods and Design Principles for Cell-Penetrant Peptides.Angewandte Chemie (International ed. in English) · 2018Review
- Nanoparticle mediated delivery and small molecule triggered activation of proteins in the nucleus.Nucleus (Austin, Tex.) · 2018Article
- A Redox-Activatable Fluorescent Sensor for the High-Throughput Quantification of Cytosolic Delivery of Macromolecules.Angewandte Chemie (International ed. in English) · 2017Article
- Mechanism Matters: A Taxonomy of Cell Penetrating Peptides.Trends in biochemical sciences · 2015Review
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Proteins and other macromolecules that cross biological membranes have great potential as tools for research and next-generation therapeutics. Here, we describe two assays that effectively quantify the cytosolic localization of a number of previously reported peptides and protein domains. One assay, which we call GIGI (glucocorticoid-induced eGFP induction), is an amplified assay that informs on relative cytosolic access without the need for sophisticated imaging equipment or adherent cells. The second, GIGT (glucocorticoid-induced eGFP translocation), is a nonamplified assay that informs on relative cytosolic access and exploits sophisticated imaging equipment to facilitate high-content screens in live cells. Each assay was employed to quantify the cytosolic delivery of several canonical "cell permeable peptides," as well as more recently reported minimally cationic miniature proteins and zinc finger nuclease domains. Our results show definitively that both overall charge as well as charge distribution influence cytosolic access and that small protein domains containing a discrete, helical, penta-Arg motif can dramatically improve the cytosolic delivery of small folded proteins such as zinc finger domains. We anticipate that the assays described herein will prove useful to explore and discover the fundamental physicochemical and genetic properties that influence both the uptake and endosomal release of peptidic molecules and their mimetics.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.