| Title : Evaluation of the Cytosolic Uptake of HaloTag Using a pH-Sensitive Dye - Giancola_2024_ACS.Chem.Biol_19_908 |
| Author(s) : Giancola JB , Grimm JB , Jun JV , Petri YD , Lavis LD , Raines RT |
| Ref : ACS Chemical Biology , 19 :908 , 2024 |
| 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. |
| ESTHER : Giancola_2024_ACS.Chem.Biol_19_908 |
| PubMedSearch : Giancola_2024_ACS.Chem.Biol_19_908 |
| PubMedID: 38525961 |
Giancola JB, Grimm JB, Jun JV, Petri YD, Lavis LD, Raines RT (2024)
Evaluation of the Cytosolic Uptake of HaloTag Using a pH-Sensitive Dye
ACS Chemical Biology
19 :908
Giancola JB, Grimm JB, Jun JV, Petri YD, Lavis LD, Raines RT (2024)
ACS Chemical Biology
19 :908