The HaloTag as a general scaffold for far-red tunable chemigenetic indicators.
Nat Chem Biol
; 17(6): 718-723, 2021 06.
Article
in En
| MEDLINE
| ID: mdl-33795886
ABSTRACT
Functional imaging using fluorescent indicators has revolutionized biology, but additional sensor scaffolds are needed to access properties such as bright, far-red emission. Here, we introduce a new platform for 'chemigenetic' fluorescent indicators, utilizing the self-labeling HaloTag protein conjugated to environmentally sensitive synthetic fluorophores. We solve a crystal structure of HaloTag bound to a rhodamine dye ligand to guide engineering efforts to modulate the dye environment. We show that fusion of HaloTag with protein sensor domains that undergo conformational changes near the bound dye results in large and rapid changes in fluorescence output. This generalizable approach affords bright, far-red calcium and voltage sensors with highly tunable photophysical and chemical properties, which can reliably detect single action potentials in cultured neurons.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fluorescent Dyes
/
Hydrolases
Limits:
Animals
Language:
En
Journal:
Nat Chem Biol
Journal subject:
BIOLOGIA
/
QUIMICA
Year:
2021
Type:
Article
Affiliation country:
United States