| Title : Engineered HaloTag variants for fluorescence lifetime multiplexing - Frei_2022_Nat.Methods_19_65 |
| Author(s) : Frei MS , Tarnawski M , Roberti MJ , Koch B , Hiblot J , Johnsson K |
| Ref : Nat Methods , 19 :65 , 2022 |
|
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. |
| PubMedSearch : Frei_2022_Nat.Methods_19_65 |
| PubMedID: 34916672 |
| Gene_locus related to this paper: rhoso-halo1 |
| Inhibitor | Chloroalkane-tetramethylrhodamine-fluorophore |
| Gene_locus | rhoso-halo1 |
| Structure | 7PCW 7PCX 6ZVY 6Y7A |
Frei MS, Tarnawski M, Roberti MJ, Koch B, Hiblot J, Johnsson K (2022)
Engineered HaloTag variants for fluorescence lifetime multiplexing
Nat Methods
19 :65
Frei MS, Tarnawski M, Roberti MJ, Koch B, Hiblot J, Johnsson K (2022)
Nat Methods
19 :65