Engineered HaloTag variants for fluorescence lifetime multiplexing.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 34916672.
- Also identified by DOI 10.1038/s41592-021-01341-x and PMC identifier 8748199.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Biosensing Techniques
- Fluorescent Dyes
- Hydrolases