为了结合这些优势，研究小组以从头开始的蛋白质设计为主题，为三种跨越可见光谱的细胞渗透性染料生成小的、纳米分子亲和的、高选择性的结合物（NovoTags）。该课题组人员展示了NovoTag荧光寿命可以调谐，并展示了它们在基于寿命和波长的多路荧光成像中的应用。课题组研究人员进一步设计了一种双链NovoTag，可作为活细胞中具有荧光读出的化学诱导二聚化系统，或作为固定细胞中最小干扰的接近探针。他们的方法结合了荧光蛋白和小分子染料的优点，扩展了细胞成像的工具箱。
研究人员表示，荧光蛋白和小分子染料为生物成像提供了互补的优势：蛋白质适合遗传标记，而染料提供优越的亮度和光稳定性。
附：英文原文
Title: De novo design of orthogonal far-red, orange, and green fluorophore-binding proteins for multiplexed imaging
Author: Long Tran, Steffen Klein, David Juergens, Shajesh Sharma, Justin Decarreau, Gyu Rie Lee, Yujia Wang, Wei Chen, Asim K. Bera, Alex Kang, Jon Woods, Emily Joyce, Dionne K Vafeados, Nicole Roullier, Xinting Li, Bingxu Liu, Yang Bo, Edin Muratspahi, Tim A. Brown, Jonathan B. Grimm, Ronak Patel, Luke D. Lavis, Julia Mahamid, Linna An, David Baker
Issue&Volume: 2026-07-16
Abstract: Fluorescent proteins and small-molecule dyes offer complementary advantages for biological imaging: proteins are amenable to genetic tagging, whereas dyes provide superior brightness and photostability. To combine these strengths, we used de novo protein design to generate small, nanomolar-affinity, high-selectivity binders (NovoTags) for three cell-permeable dyes spanning the visible spectrum. We show that the NovoTag fluorescent lifetimes can be tuned and demonstrate their application in lifetime and wavelength-based multiplexed fluorescence imaging. We further design a two-chain NovoTag that functions as a chemically induced dimerization system with fluorescent readout in living cells, or as a minimally perturbing proximity probe in fixed cells. Our approach combines the advantages of fluorescent proteins and small-molecule dyes, expanding the toolkit for cellular imaging.
DOI: aeb0822
Source: https://www.science.org/doi/10.1126/science.aeb0822