Single-domain near-infrared protein provides a scaffold for antigen-dependent fluorescent nanobodies

被引:0
作者
Olena S. Oliinyk
Mikhail Baloban
Charles L. Clark
Erin Carey
Sergei Pletnev
Axel Nimmerjahn
Vladislav V. Verkhusha
机构
[1] University of Helsinki,Medicum, Faculty of Medicine
[2] Albert Einstein College of Medicine,Department of Genetics and Gruss
[3] Salk Institute for Biological Studies,Lipper Biophotonics Center
[4] National Institutes of Health,Waitt Advanced Biophotonics Center
来源
Nature Methods | 2022年 / 19卷
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摘要
Small near-infrared (NIR) fluorescent proteins (FPs) are much needed as protein tags for imaging applications. We developed a 17 kDa NIR FP, called miRFP670nano3, which brightly fluoresces in mammalian cells and enables deep-brain imaging. By exploring miRFP670nano3 as an internal tag, we engineered 32 kDa NIR fluorescent nanobodies, termed NIR-Fbs, whose stability and fluorescence strongly depend on the presence of specific intracellular antigens. NIR-Fbs allowed background-free visualization of endogenous proteins, detection of viral antigens, labeling of cells expressing target molecules and identification of double-positive cell populations with bispecific NIR-Fbs against two antigens. Applying NIR-Fbs as destabilizing fusion partners, we developed molecular tools for directed degradation of targeted proteins, controllable protein expression and modulation of enzymatic activities. Altogether, NIR-Fbs enable the detection and manipulation of a variety of cellular processes based on the intracellular protein profile.
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页码:740 / 750
页数:10
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