Engineered HaloTag variants for fluorescence lifetime multiplexing

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Frei, Michelle S., Tarnawski, Miroslaw, Roberti, M. Julia, Koch, Birgit, Hiblot, Julien, Johnsson, Kai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group US 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8748199/
https://www.ncbi.nlm.nih.gov/pubmed/34916672
http://dx.doi.org/10.1038/s41592-021-01341-x
Descripción
Sumario: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.

Ejemplares similares