ArticleACS chemical biology2024
Evaluation of the Cytosolic Uptake of HaloTag Using a pH-Sensitive Dye.
Article in ACS chemical biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Spiro-helicenoid Carbofluoresceins: CPL-Active Emitters for Bioimaging.Organic letters · 2026Article
- Protein Modifications for Cellular Protein Delivery.Chemical reviews · 2026Review
- Membrane-Traversing Peptides: Mechanistic Landscapes, Assays, and Applications.Chemical reviews · 2026Review
- Hydrophilic α-Aryl-α-Diazoamides for Protein Esterification.Synlett : accounts and rapid communications in synthetic organic chemistry · 2025Article
- Cytosolic Delivery of Functional Ubiquitin.Journal of peptide science : an official publication of the European Peptide Society · 2025Article
- Sculpting excitable membranes: voltage-gated ion channel delivery and distribution.Nature reviews. Neuroscience · 2025Review
- Mammalian Esterase Activity: Implications for Peptide Prodrugs.Biochemistry · 2024Article
- Regulation of Absorption and Emission in a Protein/Fluorophore Complex.ACS chemical biology · 2024Article
- Facile Access to Branched Multispecific Proteins.Bioconjugate chemistry · 2024Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
The efficient cytosolic delivery of proteins is critical for advancing novel therapeutic strategies. Current delivery methods are severely limited by endosomal entrapment, and detection methods lack sophistication in tracking the fate of delivered protein cargo. HaloTag, a commonly used protein in chemical biology and a challenging delivery target, is an exceptional model system for understanding and exploiting cellular delivery. Here, we employed a combinatorial strategy to direct HaloTag to the cytosol. We established the use of Virginia Orange, a pH-sensitive fluorophore, and Janelia Fluor 585, a similar but pH-agnostic fluorophore, in a fluorogenic assay to ascertain protein localization within human cells. Using this assay, we investigated HaloTag delivery upon modification with cell-penetrating peptides, carboxyl group esterification, and cotreatment with an endosomolytic agent. We found efficacious cytosolic entry with two distinct delivery methods. This study expands the toolkit for detecting the cytosolic access of proteins and highlights that multiple intracellular delivery strategies can be used synergistically to effect cytosolic access. Moreover, HaloTag is poised to serve as a platform for the delivery of varied cargo into human cells.
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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.