小柯机器人

德国马克斯-普朗克医学研究所Kai Johnsson小组开发出工程化的HaloTag变体用于荧光寿命复用。该研究于2021年12月16日在线发表于国际一流学术期刊《自然—方法学》。

研究人员介绍了HaloTag的变体，与HaloTag7相比，当用罗丹明标记时，HaloTag的亮度和荧光寿命增加或减少。结合这些HaloTag变体，可以在一个光谱通道中使用单一荧光团对三个细胞靶标进行活体荧光寿命复用，并产生一个基于荧光寿命的生物传感器。此外，最亮的HaloTag变体在活细胞成像应用中显示了高达40%的亮度。

据悉，像HaloTag这样的自标记蛋白标签是强大的工具，可以用合成的荧光团标记融合蛋白，用于荧光显微镜使用。

附：英文原文

Title: Engineered HaloTag variants for fluorescence lifetime multiplexing

Author: Frei, Michelle S., Tarnawski, Miroslaw, Roberti, M. Julia, Koch, Birgit, Hiblot, Julien, Johnsson, Kai

Issue&Volume: 2021-12-16

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. HaloTag variants offer distinct brightness and fluorescence lifetimes compared with HaloTag7 when labeled with rhodamines. These variants were used for multiplexed imaging with a single fluorophore and to create lifetime-based cell cycle indicators.

DOI: 10.1038/s41592-021-01341-x

Source: https://www.nature.com/articles/s41592-021-01341-x

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex