Engineered HaloTag variants for fluorescence lifetime multiplexing.
Nat Methods
; 19(1): 65-70, 2022 01.
Article
em En
| MEDLINE
| ID: mdl-34916672
ABSTRACT
Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce HaloTag variants with either increased or decreased brightness and fluorescence lifetime compared with HaloTag7 when labeled with rhodamines. Combining these HaloTag variants enabled live-cell fluorescence lifetime multiplexing of three cellular targets in one spectral channel using a single fluorophore and the generation of a fluorescence lifetime-based biosensor. Additionally, the brightest HaloTag variant showed up to 40% higher brightness in live-cell imaging applications.
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1
Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
/
Corantes Fluorescentes
/
Hidrolases
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article